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The exterior of bacteriophage T4 capsid is coated with two outer capsid proteins, Hoc (highly antigenic outer capsid protein; molecular mass, 40 kDa) and Soc (small outer capsid protein; molecular mass, 9 kDa), at symmetrical positions on the icosahedron (160 copies of Hoc and 960 copies of Soc per capsid particle). Both these proteins are nonessential for phage infectivity and viability and assemble onto the capsid surface after completion of capsid assembly. We developed a phage display system which allowed in-frame fusions of foreign DNA at a unique cloning site in the 5 end of hoc or soc. A DNA fragment corresponding to the 36-amino-acid PorA peptide from Neisseria meningitidis was cloned into the display vectors to generate fusions at the N terminus of Hoc or Soc. The PorA-Hoc and PorA-Soc fusion proteins retained the ability to bind to the capsid surface, and the bound peptide was displayed in an accessible form as shown by its reactivity with specific monoclonal antibodies in an ...

TY - JOUR. T1 - The herpesvirus capsid surface protein, VP26, and the majority of the tegument proteins are dispensable for capsid transport toward the nucleus. AU - Antinone, Sarah E.. AU - Shubeita, George T.. AU - Coller, Kelly E.. AU - Lee, Joy I.. AU - Haverlock-Moyns, Sarah. AU - Gross, Steven P.. AU - Smith, Gregory A.. N1 - Copyright: Copyright 2011 Elsevier B.V., All rights reserved.. PY - 2006/6. Y1 - 2006/6. N2 - Upon entering a cell, alphaherpesvirus capsids are transported toward the minus ends of microtubules and ultimately deposit virus DNA within the host nucleus. The virus proteins that mediate this centripetal transport are unknown but are expected to be either viral tegument proteins, which are a group of capsid-associated proteins, or a surface component of the capsid itself. Starting with derivatives of pseudorabies virus that encode a fluorescent protein fused to a structural component of the virus, we have made a collection of 12 mutant viruses that lack either the VP26 ...

Retroviral capsid proteins are conserved structurally but assemble into different morphologies. The mature human immunodeficiency virus-1 (HIV-1) capsid is best described by a fullerene cone model, in which hexamers of the capsid protein are linked to form a hexagonal surface lattice that is closed by incorporating 12 capsid-protein pentamers. HIV-1 capsid protein contains an amino-terminal domain (NTD) comprising seven α-helices and a β-hairpin, a carboxy-terminal domain (CTD) comprising four α-helices, and a flexible linker with a 310-helix connecting the two structural domains. Structures of the capsid-protein assembly units have been determined by X-ray crystallography; however, structural information regarding the assembled capsid and the contacts between the assembly units is incomplete. Here we report the cryo-electron microscopy structure of a tubular HIV-1 capsid-protein assembly at 8 Å resolution and the three-dimensional structure of a native HIV-1 core by cryo-electron tomography. The

Post-translational modifications in viral capsids are known to fine-tune and regulate several aspects of the infective life cycle of several viruses in the host. Recombinant viruses that are generated in a specific producer cell line are likely to inherit unique post-translational modifications during intra-cellular maturation of its capsid proteins. Data on such post-translational modifications in the capsid of recombinant adeno-associated virus serotypes (AAV1-rh10) is limited. We have employed liquid chromatography and mass spectrometry analysis to characterize post-translational modifications in AAV1-rh10 capsid protein. Our analysis revealed a total of 52 post-translational modifications in AAV2-AAVrh10 capsids, including ubiquitination (17%), glycosylation (36%), phosphorylation (21%), SUMOylation (13%) and acetylation (11%). While AAV1 had no detectable post-translational modification, at least four AAV serotypes had ,7 post-translational modifications in their capsid protein. About 82% ...

Author summary The picornavirus family includes clinically important human and animal pathogens, for example: poliovirus, rhinovirus (causing the common cold) and foot-and-mouth disease virus (FMDV) that infects cloven-hoofed animals. Picornaviruses contain a positive-sense RNA genome surrounded by a protein shell, also called a capsid. The capsid proteins are made from a precursor and correct processing and assembly of these capsid proteins is necessary in the virus life cycle to create new infectious virus particles. In this study, we have identified a short motif (just 5 amino acids long) within the capsid precursor, which is highly conserved among picornaviruses. Deletion of this motif inhibited processing of the junctions between the mature structural proteins within this precursor, with one junction being more than 400 amino acids away from this region. This motif also seems to be required for the optimal accumulation of the capsid precursor in cells. We hypothesize that the motif may be involved

Viral capsids are metastable structures that perform many essential processes; they also act as robust cages during the extracellular phase. Viruses can use multifunctional proteins to optimize resources (e.g., VP3 in avian infectious bursal disease virus, IBDV). The IBDV genome is organized as ribonucleoproteins (RNP) of dsRNA with VP3, which also acts as a scaffold during capsid assembly. We characterized mechanical properties of IBDV populations with different RNP content (ranging from none to four RNP). The IBDV population with the greatest RNP number (and best fitness) showed greatest capsid rigidity. When bound to dsRNA, VP3 reinforces virus stiffness. These contacts involve interactions with capsid structural subunits that differ from the initial interactions during capsid assembly. Our results suggest that RNP dimers are the basic stabilization units of the virion, provide better understanding of multifunctional proteins, and highlight the duality of RNP as capsid-stabilizing and genetic ...

Herpesvirus capsid protein. Molecular model of the major capsid protein VP5 from herpes simplex virus-1 (HSV-1). This virus infects epithelial cells around the mouth, causing fluid-filled blisters known as cold sores. There is no known cure for herpes infection and the virus remains in the body for life. - Stock Image F006/9435

Summary The inner capsid structure of the OSU strain of porcine rotavirus was studied by electron microscopy of freeze-dried preparations and of negatively stained chemically disrupted virus particles. The analysis of the particles by the freeze-drying technique revealed a T:13 l (laevo) symmetry for the organization of the inner capsid. Treatment of single-capsid rotavirus particles with 30% formamide or 5 m-urea resulted in their degradation, giving rise to very similar products, corresponding to isolated vertices, edges and faces of the virus icosahedron. An analysis of such structures confirmed the triangulation number and handedness of the rotavirus inner capsid, and provided evidence for the open-mesh model, in which the five- and six-coordinated axes are represented by holes formed by smaller trimeric morphological subunits.

TY - JOUR. T1 - The herpesvirus VP1/2 protein is an effector of dynein-mediated capsid transport and neuroinvasion. AU - Zaichick, Sofia V.. AU - Bohannon, Kevin P.. AU - Hughes, Ami. AU - Sollars, Patricia J.. AU - Pickard, Gary E.. AU - Smith, Gregory A.. PY - 2013/2/13. Y1 - 2013/2/13. N2 - Microtubule transport of herpesvirus capsids from the cell periphery to the nucleus is imperative for viral replication and, in the case of many alphaherpesviruses, transmission into the nervous system. Using the neuroinvasive herpesvirus, pseudorabies virus (PRV), we show that the viral protein 1/2 (VP1/2) tegument protein associates with the dynein/dynactin microtubule motor complex and promotes retrograde microtubule transport of PRV capsids. Functional activation of VP1/2 requires binding to the capsid protein pUL25 or removal of the capsid-binding domain. A proline-rich sequence within VP1/2 is required for the efficient interaction with the dynein/dynactin microtubule motor complex as well as for PRV ...

The crystal structure of the double-stranded DNA bacteriophage HK97 mature empty capsid was determined at 3.6 angstrom resolution. The 660 angstrom diameter icosahedral particle contains 420 subunits with a new fold. The final capsid maturation step is an autocatalytic reaction that creates 420 isopeptide bonds between proteins. Each subunit is joined to two of its neighbors by ligation of the side-chain lysine 169 to asparagine 356. This generates 12 pentameric and 60 hexameric rings of covalently joined subunits that loop through each other, creating protein chainmail: topologically linked protein catenanes arranged with icosahedral symmetry. Catenanes have not been previously observed in proteins and provide a stabilization mechanism for the very thin HK97 capsid. ...

Adeno-associated virus capsid, molecular model. The capsid is a protein shell that encloses the virus genetic information. Adeno-associated viruses (AAVs) are the smallest known viruses to infect humans. This is serotype 6 of the virus. AAVs do not cause diseases, and only provoke a mild immune response. Because they incorporate their genetic material into a specific location within the hosts genome, they have potential as a vector for gene therapy. - Stock Image C014/2836

TY - JOUR. T1 - Partial amino-terminal sequences of the polyoma nonhistone proteins VP1, VP2, and VP3 synthesized in vitro. AU - Hewick, R. M.. AU - Mellor, A.. AU - Smith, A. E.. AU - Waterfield, M. D.. N1 - Copyright: Copyright 2020 Elsevier B.V., All rights reserved.. PY - 1980. Y1 - 1980. N2 - The three polyoma virus capsid proteins VP1, VP2, and VP3 were synthesized in vitro in the presence of several radiolabeled amino acids and, after purification on sodium dodecyl sulfate-polyacrylamide gels, were subjected to sequential Edman degradation. The partial amino-terminal amino acid sequences obtained were compared with the sequence of amino acids predicted from the polyoma virus DNA sequencing. Together, these results showed that the 5 ends of the VP1, VP2, and VP3 coding sequences are located 1,217, 289, and 634 nucleotides, respectively, from the junction of HpaII restriction fragments 3 and 5.. AB - The three polyoma virus capsid proteins VP1, VP2, and VP3 were synthesized in vitro in the ...

Adenoviral vectors have great potential for use in gene therapy and genetic immunization. The targeting of Ad vectors to the relevant tissue and cell types in vivo could greatly improve their safety and performance by lowering the effective dosage required for therapeutic levels of gene expression. Redirection of Ad vector tropism will require physical modifications of the adenoviral capsid but direct genetic modification of the Ad capsid has so far been limited to small peptides. A novel system for the attachment of targeting ligands to the Ad capsid, based on the extremely strong avidin-biotin interaction, is described herein. The genetic insertion of a biotin acceptor peptide (BAP) into the fiber, protein IX, or hexon components of the Ad capsid has resulted in vectors that are metabolically biotinylated upon production in host cells. Avidin-dependent redirection of transduction through a variety of biotinylated ligands is greatly dependent on the nature of the biotinylated capsid protein. ...

Lien vers Pubmed [PMID] - 21512573. Nature 2011 Apr;472(7343):361-5. TRIM5 is a RING domain-E3 ubiquitin ligase that restricts infection by human immunodeficiency virus (HIV)-1 and other retroviruses immediately following virus invasion of the target cell cytoplasm. Antiviral potency correlates with TRIM5 avidity for the retrovirion capsid lattice and several reports indicate that TRIM5 has a role in signal transduction, but the precise mechanism of restriction is unknown. Here we demonstrate that TRIM5 promotes innate immune signalling and that this activity is amplified by retroviral infection and interaction with the capsid lattice. Acting with the heterodimeric, ubiquitin-conjugating enzyme UBC13-UEV1A (also known as UBE2N-UBE2V1), TRIM5 catalyses the synthesis of unattached K63-linked ubiquitin chains that activate the TAK1 (also known as MAP3K7) kinase complex and stimulate AP-1 and NFκB signalling. Interaction with the HIV-1 capsid lattice greatly enhances the UBC13-UEV1A-dependent E3 ...

Human papilloma viruses (HPV) are small DNA, non-enveloped, double-stranded and closed circular viruses. There are more than 150 HPV identified types. Genital HPV types are categorized according to their epidemiologic association with cervical cancer to high and low risk types. The high risk type HPV 16 is the most common in the world. we aimed to design a universal peptide based vaccine against HPV type 16 virus using Immunoinformatics Approach through prediction of highly conserved T and B-cell epitopes from the most abundant and highly immunogenic protein (L1 major capsid protein) derived from HPV type 16 strains all over the world. All sequences of the L1 major capsid protein were retrieved from NCBI database. Potentially continuous B and T cell epitopes were predicted using tools from immune epitope data base analysis resource (IEDB-AR). The Allergenicity of predicted epitopes was analyzed by AllerTOP Tool and the coverage was determined throughout the worlds. The B cell epitope 243KSEV246 ...

Assembly of recombinant capsid proteins into virus-like particles (VLPs) still represents an interesting challenge in virus-based nanotechnologies. The structure of VLPs has gained importance for the development and design of new adjuvants and antigen carriers. The potential of Tobacco etch virus capsid protein (TEV CP) as adjuvant has not been evaluated to date. Two constructs for TEV CP expression in Escherichia coli were generated: a wild-type version (TEV-CP) and a C-terminal hexahistidine (His)-tagged version (His-TEV-CP). Although both versions were expressed in the soluble fraction of E. coli lysates, only His-TEV-CP self-assembled into micrometric flexuous filamentous VLPs. In addition, the His-tag enabled high yields and facilitated purification of TEV VLPs. These TEV VLPs elicited broader IgG2-specific antibody response against a novel porcine reproductive and respiratory syndrome virus (PRRSV) protein when compared to the potent IgG1 response induced by the protein alone. His-TEV CP was

Foamy viruses (FVs) of the Spumaretrovirinae subfamily are distinct retroviruses, with many features of their molecular biology and replication strategy clearly different from those of the Orthoretroviruses, such as human immunodeficiency, murine leukemia, and human T cell lymphotropic viruses. The FV Gag N-terminal region is responsible for capsid formation and particle budding via interaction with Env. However, the critical residues or motifs in this region and their functional interaction are currently ill-defined, especially in non-primate FVs. Mutagenesis of N-terminal Gag residues of feline FV (FFV) reveals key residues essential for either capsid assembly and/or viral budding via interaction with the FFV Env leader protein (Elp). In an in vitro Gag-Elp interaction screen, Gag mutations abolishing particle assembly also interfered with Elp binding, indicating that Gag assembly is a prerequisite for this highly specific interaction. Gradient sedimentation analyses of cytosolic proteins indicate

Capsids of the B19 parvovirus are composed of major (VP2; 58 kD) and minor (VP1; 83 kD) structural proteins. These proteins are identical except for a unique 226 amino acid region at the amino terminus of VP1. Previous immunization studies with recombinant empty capsids have demonstrated that the presence of VP1 was required to elicit virus-neutralizing antibody activity. However, to date, neutralizing epitopes have been identified only on VP2. Crystallographic studies of a related parvovirus (canine parvovirus) suggested the unique amino-terminal portion of VP1 assumed an internal position within the viral capsid. To determine the position of VP1 in both empty capsids and virions, we expressed a fusion protein containing the unique region of VP1. Antisera raised to this protein recognized recombinant empty capsids containing VP1 and VP2, but not those containing VP2 alone, in an enzyme-linked immunosorbent assay. The antisera immunoprecipitated both recombinant empty capsids and human ...

Samuel Campos, PhD, studies early events of Human Papillomavirus (HPV) infection. HPVs are small, non-enveloped DNA viruses that cause a variety of lesions ranging from benign waters to cervical cancers. Although over 100 types of HPVs have been identified, HPV16 is the most prevalent, and is alone responsible for more than 50% of cervical cancers in women worldwide. Dr. Campos and his lab study the mechanisms of HPV virus transmission at a cellular level, in hopes to discover new approaches for the prevention and treatment of HPV.HPV16 virions consist of an ~8kb circular dsDNA genome packaged into a ~60 nm protein capsid. The genome is condensed with cellular histones and exists in a chromatin-like state. The capsid is comprised of 72 pentamers of the major capsid protein L1 and up to 72 molecules of the minor capsid protein L2, localized along the inner capsid surface, within the central cavities beneath the L1 pentamers. Mature HPV16 virions exist in an oxidized state, with adjacent L1 ...

In a mature and infectious retroviral particle, the capsid protein (CA) forms a shell surrounding the genomic RNA and the replicative machinery of the virus. The irregular nature of this capsid shell precludes direct atomic resolution structural analysis. CA hexamers and pentamers are the fundamental building blocks of the capsid, however the pentameric state, in particular, remains poorly characterized. We have developed an efficient in vitro protocol for studying the assembly of Rous sarcoma virus (RSV) CA that involves mild acidification and produces structures modeling the authentic viral capsid. These structures include regular spherical particles with T = 1 icosahedral symmetry, built from CA pentamers alone. These particles were subject to cryoelectron microscopy (cryo-EM) and image processing, and a pseudo-atomic model of the icosahedron was created by docking atomic structures of the constituent CA domains into the cryo-EM-derived three-dimensional density map. The N-terminal domain ...

One monoclonal antibody (mAb; 8E4), against the capsid protein of PCV2, was generated and characterised in this study. 8E4 reacted with the genotype PCV2a (CL, LG and JF2) strains but not PCV2b (YJ, SH and JF) strains by an immunoperoxidase mono-layer assay (IPMA) and a capture ELISA. Furthermore, the mAb had the capacity to neutralise PCV2a (CL, LG and JF2) strains but not PCV2b (YJ, SH and JF) strains. One critical amino acid that determined a conformational neutralising epitope was identified using mAb 8E4 and PCV2 infectious clone technique. Amino acid residues 47-72 in the capsid protein of PCV2a/CL were replaced with the corresponding region of PCV2b/YJ, and the reactivity of mAb 8E4 was lost. Further experiments demonstrated that one amino acid substitution, the alanine for arginine at position 59 (A59R) in the capsid protein of PCV2a (CL, LG and JF2) strains, inhibited completely the immunoreactivity of three PCV2a strains with mAb 8E4 ...

Due to the diversity of their biological properties, the serotypes of AAV are important resources for the gene therapy community. In order to develop the full potential of AAV vectors for gene therapy applications, it is of interest to define the capsid domains of the individual serotypes responsible for these properties. The ability to interchange these regions should facilitate designing AAV vectors with novel tropisms and transduction properties.. Domain swapping via various cloning methods as a means to exchange serotype specific properties such as receptor binding, tissue tropism, and purification schemes has been proposed (30). Xiao et al. identified regions of the AAV1 capsid responsible for its ability to transduce skeletal muscle by systematically exchanging domains between AAV1 and AAV2. This approach, which relied on cloning and assessing each individual construct, was successful and demonstrates the importance of such a strategy (W. Xiao et al., submitted for publication).. We used a ...

PubMed journal article: High prevalence of immunoglobulin A antibody against Epstein-Barr virus capsid antigen in adult patients with lupus with disease flare: case control studies. Download Prime PubMed App to iPhone, iPad, or Android

Capsid vertex-specific component that plays a role during viral DNA encapsidation, assuring correct genome cleavage and presumably stabilizing capsids that contain full-length viral genomes. Participates in the interaction between the capsid and the tegument through interaction with the large tegument protein/LTP.

Our main goal was to produce a library of Adeno-associated viruses that infect cells with different specificities and efficiencies, due to differing capsid composition, and select from this library the best AAVs that are highly specific. Since it is not the case that viruses that work well in vitro will work as good in vivo, we had the aim of testing our candidates in mice as well. To achieve this, the capsid genes from AAV serotypes 1,2,5,6,8 and 9 were shuffled between each other in a primerless PCR reaction that relies on the close homology between the different cap genes. Those capsid genes were first digested with DNase I, then pooled together and allowed to anneal to each other. A second PCR was done then to amplify the shuffled cap gene fragments that were generated in the first PCR while introducing AscI and PacI restriction sites, which are used to clone the cap genes into a helper vector. AAVs that were then produced in HEK293 cells, which were transfected with the cap-gene-ITR ...

Singararn SW, Garmann RF, Knobler CM, Gelbart WM, Ben-Shaul A. Role of RNA Branchedness in the Competition for Viral Capsid Proteins. JOURNAL OF PHYSICAL CHEMISTRY B. 2015;119 :13991-14002.

The individual papillomavirus type 11 (HPV-11) L1 main capsid protein could be trypsinized to create recombinant capsomeres that retain HPV genotype-restricted capsid antigenicity (M. immunogenic highly, genotype-restricted HPV capsid-neutralizing antigenic domains are included within capsomeres entirely. Thus, capsomeres may be viable vaccine applicants for preventing HPV disease. Papillomaviruses trigger hyperproliferative cutaneous and/or mucosal epithelial lesions in higher vertebrates, including human beings (31). A lot more than 70 genotypically specific human being papillomaviruses (HPVs) have already been identified (12) and may be categorized based on observed variations in disease phenotype (i.e., desired cells tropisms and/or disease organizations). For instance, most HPVs preferentially infect cutaneous pores and skin and usually trigger only harmless disease (e.g., plantar or common warts), while other styles even more infect PF 477736 oral or anogenital mucosal epithelium often. ...

It is becoming increasingly clear that mammalian cells can express a variety of factors that limit retroviral replication. One class of these factors blocks replication at a stage post-entry but prior to nuclear import and integration of the viral genome. Members of this family arc known as restriction factors. The best characterised member is the mouse gene hvl which determines susceptibility to infection by MLV. I-vl has two alleles known as Fvl and Fv/ whose restriction characteristics determine the host range of different MLV strains. It has become apparent that the phenomenon of restriction is not limited to MLV and murine cells. HIV-1, as well as MLV is restricted at a post-entry step in a number of primate species including rhesus macaques. African green monkeys and owl monkeys. The Thm5 gene in these species is the main determinant of both HIV-1 and MLV restriction. Viral determinants of this restriction map to the virus capsid protein, however despite strong genetic ev idence, no ...

Hepatitis B virus (HBV) infections represent a significant burden on global public health. Current HBV treatments using nucleos(t)ide analogs (NAs) and PEG interferons cannot fully alleviate this burden as they do not affect the transcriptional activity of the tenacious covalently closed circular DNA (cccDNA) responsible for viral persistence. Capsid assembly modulators (CAMs) disrupt the encapsidation of pre-genomic RNA and can cause nucleocapsid disassembly, thereby affecting multiple steps of HBV replication and reduction of cccDNA pools. This review provides a concise overview of the development of CAMs and the progress achieved in understanding their interactions with HBV core proteins. ...

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Nevertheless, up to now, this passive immune technique has not been carried out as a therapy for RVA-linked diarrhea. Although Ab muscles can have higher

The L1 major capsid proteins of human papillomavirus (HPV) types 11 and 16 were purified and analyzed for structural integrity and in vitro self-assembly. Proteins were expressed in Escherichia coli as glutathione-S-transferase-L1 (GST-L1) fusions and purified to near homogeneity as pentamers (equiv …

What Additional Protective Layer Of Protein Surrounds Some Viruses - The capsid and envelope play many roles in viral contamination, inclusive of virus attachment to cells, entry into cells, release of the capsid contents into the cells, and packaging of newly fashioned viral debris. The capsid and envelope are also answerable for transfer of the viral genetic material from one mobile to another. these systems also decide the stableness traits of the virus particle, which includes resistance to chemical or bodily inactivation.. The primary role of capsid is to bundle the viral genome. There are strategies related to this feature: the recruitment of the viral RNA all through assembly and the discharge of the genome at some stage in infection. even though particle assembly takes vicinity on endoplasmic reticulum membranes, capsid localizes in nucleoli and lipid droplets.. Because the capsid is outermost, most agents of inactivation must act on the capsid before they can reach the viral RNA, and in ...

EBV Antibody To Viral Capsid Antigen, IgM,ARUP Laboratories is a national reference laboratory and a worldwide leader in innovative laboratory research and development. ARUP offers an extensive test menu of highly complex and unique medical tests in clinical and anatomic pathology. Owned by the University of Utah, ARUP Laboratories client,medicine,medical supply,medical supplies,medical product

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Minor protein of the capsid that localizes along the inner surface of the virion, within the central cavities beneath the L1 pentamers. Plays a role in capsid stabilization through interaction with the major capsid protein L1. Once the virion enters the host cell, L2 escorts the genomic DNA into the nucleus by promoting escape from the endosomal compartments and traffic through the host Golgi network. Mechanistically, the C-terminus of L2 possesses a cell-penetrating peptide that protudes from the host endosome, interacts with host cytoplasmic retromer cargo and thereby mediates the capsid delivery to the host trans-Golgi network. Plays a role through its interaction with host dynein in the intracellular microtubule-dependent transport of viral capsid toward the nucleus. Mediates the viral genome import into the nucleus through binding to host importins. Once within the nucleus, L2 localizes viral genomes to host PML bodies in order to activate early gene expression for establishment of ...

Under conditions of low ionic strength and a pH ranging between about 3.7 and 5.0,solutions of purified coat proteins of cowpea chlorotic mottle virus (CCMV) form sphericalmultishell structures in the absence of viral RNA. The outer surfaces of the shells in thesestructures are negatively charged, whereas the inner surfaces are positively charged due to a disordered cationic N-terminal domain of the capsid protein, the arginine-rich RNA-binding motif that protrudes into the interior. We show that the main forces stabilizing these .... ...

The structure of bacteriophage SPP1 capsid was determined at sub-nanometer resolution by cryo-electron microscopy and single-particle analysis. The icosahedral capsid is composed of the major capsid protein gp13 and the auxiliary protein gp12 that are organized in a T=7 lattice. DNA is arranged in layers with a distance of ∼24.5 Å. Gp12 forms spikes that are anchored at the centre of gp13 hexamers. In a gp12-deficient mutant the centres of hexamers are closed by loops of gp13 coming together to protect the SPP1 genome from the outside environment. The HK97-like fold was used to build a pseudo-atomic model of gp13. Its structural organization remains unchanged upon tail binding and following DNA release. Gp13 exhibits enhanced thermostability in the DNA-filled capsid. A remarkable convergence between thermostability of the capsid and of the other virion components was found, revealing that the overall architecture of the SPP1 infectious particle co-evolved towards high robustness.. ...

ViroBytes is a modified BioBytes procedure for rational shuffling of capsid genes from natural isolates of Adeno-Associated Virus. Main motivation for the new protocol is unsatisfactory incorporation of certain serotypes (and especially certain parts of AAV capsid regions eg. from AAV5) using conventional shuffling strategies (Grimm et al., 2008). Application of magnetic beads for controlled assembly and the principle of BioByte formation and annealing persists. Sticky overhangs are used for selective combination of the bytes but different method is used for the production of individual ViroBytes. AAV serotypes 1,2,5,6,8 and 9 were selected as suitable candidates for fragmentation due to their exceptional individual properties. The analysis of Cap gene sequences revealed multiple homology regions which were then used for rational fragment formation. Total number of fragments per Cap gene is eight in our case and all fragments have similar length around ~250bp to assure similar behaviour in the ...

Uppsala University, Teknisk-naturvetenskapliga vetenskapsområdet, Faculty of Science and Technology, Biology, Department of Cell and Molecular Biology. Uppsala University, Teknisk-naturvetenskapliga vetenskapsområdet, Faculty of Science and Technology, Biology, Department of Cell and Molecular Biology, Structural Molecular Biology. STRUCTURAL MOLECULAR BIOLOGY. ...

Defensins are effectors of the innate immune response with potent antibacterial activity. Their role in antiviral immunity, particularly for non-enveloped viruses, is poorly understood. We recently found that human alpha-defensins inhibit human adenovirus (HAdV) by preventing virus uncoating and rel …

Capsid protein that self-associates to form 11 pentons, building the T=13 laevo capsid in association with 160 hexamers of gp23* and one dodecamer of gp20.

The viral genome is packaged in a regular icosahedral protein shell (capsid) composed of 162 capsomeres (see Fig. 214e-1). The outer covering of the virus is a lipid-containing membrane (envelope) acquired as the DNA-containing capsid buds through the inner nuclear membrane of the host cell. Between the capsid and lipid bilayer of the envelope is the tegument. Viral replication has both nuclear and cytoplasmic phases. Initial attachment to the cell membrane involves interactions of viral glycoproteins C and B with several cellular heparan sulfate-like surface receptors. Subsequently, viral glycoprotein D binds to cellular co-receptors that belong to the tumor necrosis factor receptor family of proteins, the immunoglobulin superfamily (nectin family), or both. The ubiquity of these receptors contributes to the wide host range of herpesviruses. HSV replication is highly regulated. After fusion and entry, the nucleocapsid enters the cytoplasm and several viral proteins are released from the virion. ...

The virions of Herpesvirales order have complex and characteristic structures consisting of both symmetrical and nonsymmetrical components. The spherical virion is composed of the core, capsid, tegument and envelope. The core consists of the viral genome packaged as a single, linear, dsDNA molecule into a preformed capsid and the DNA is packed in a liquid-crystalline array that fills the entire internal volume of the capsid. The capsids assemble by cocondensation around a protein scaffold to form a procapsid in which the subunits are weakly connected. Proteolytic cleavage of the scaffolding protein triggers loss of scaffold and reorganization of the shell into the characteristic capsid form. The structure of the tegument is poorly defined, with evidence of symmetry only in the region immediately adjacent to the capsid. The tegument contains various proteins, not all of which are required for the formation of virions. The genomes are composed of linear dsDNA ranging from 125 to 295 kbp in size and from

Phage display is a method to discover peptide ligands while minimizing and optimizing the structure and function of proteins (Hallahan, 2003). The phage is used as a scaffold to display recombinant libraries of peptides and provides a means to recover and amplify the peptides that bind to putative receptor molecules in vivo. In vivo selection simultaneously provides positive and subtractive screens because organs and tissues such as tumors are spatially separated. Phage DNA can then be sequenced to determine the amino acid sequence of peptides on the capsid that have been recovered from tumors. The T7 phage display system exploits the T7 capsid protein as a scaffold to display peptides on the capsid protein unique to the 10B protein on the surface of the phage. Gene 10 encoding the capsid protein is cloned with a series of multiple cloning sites at the C-terminus of the 10B protein. The natural translational frame shift site within the capsid gene has been removed so that only a single form of ...

Video, Wyss researchers have created a high-throughput platform to generate an Adeno-associated virus 2 (AAV2) library containing 200,000 variants, each carrying a distinct mutation in the virus capsid protein.

TY - JOUR. T1 - Turnip yellow mosaic virus and its capsid have thermal stabilities with opposite ph dependence. T2 - studies by differential scanning calorimetry and 31P nuclear magnetic resonance spectroscopy. AU - Virudachalam, R.. AU - Low, Philip S.. AU - Argos, Patrick. AU - Markley, John L.. PY - 1985/10/30. Y1 - 1985/10/30. N2 - In the differential scanning calorimetry (DSC) scans of turnip yellow mosaic virus (TYMV) or its capsid a single endotherm was observed. The endotherm was attributed to disruption of the virion or capsid structure with accompanying protein denaturation. At pH 4.5 the thermal stabilities of the TYMV virion and capsid were similar. With increasing pH, the capsid stability increased while the virion stability decreased. At neutral pH the capsid disrupted at 83.5°, and the virion disrupted at 69°. Our results suggest that packaging of viral RNA in the TYMV capsid imparts instability. The pHmid for disruption of the TYMV capsid is 5.7, which is in the pKa range ...

Summary Direct evidence linking the capsid protein to specific dsRNA segments from the three killer strains of Ustilago maydis virus (P1, P4, P6) is presented. The capsid proteins of the three strains cross-react immunologically, have similar mol. wt. and similar peptide maps after limited proteolysis. The capsid proteins from P1 and P4 were translated from their respective H2 dsRNA segments, whereas the capsid protein for P6 was translated from H1 dsRNA. These in vitro translation products were each precipitated by the antiserum to capsid proteins of all three strains, had similar mol. wt. and similar peptide maps. All in vitro translation products competed effectively with native capsid proteins of all of the three strains in immunocompetition assays. These results suggest that the three strains code for a similar capsid protein, and that the information for capsid protein resides in the H2 segment of strain P1 and P4, and in the H1 segment of strain P6.

TY - THES. T1 - Chemical virology. T2 - decorating the interior of the cowpea chlorotic mottle virus. AU - Minten, Inge Jeannette. PY - 2011/4/4. Y1 - 2011/4/4. KW - METIS-283479. M3 - PhD Thesis - Research external, graduation external. SN - 978-90--9025926-0. PB - Radboud University Nijmegen. CY - Nijmegen. ER - ...

The Epstein-Barr Virus Capsid Antigen (VCA) IgM ELISA Kit is intended for the measurement of IgM antibodies to Epstein-Barr Virus Capsid Antigen (VCA) in a sample. This kit utilizes Epstein-Barr VCA antigen (P3H3 cell extract, cultured in human Burkitt lymphoma cells).

Abstract: The nuclear phase of herpesvirus replication is regulated through the formation of regulatory multi-component protein complexes. Viral genomic replication is followed by nuclear capsid assembly, DNA encapsidation and nuclear egress. The latter has been studied intensely pointing to the formation of a viral core nuclear egress complex (NEC) that recruits a multimeric assembly of viral and cellular factors for the reorganization of the nuclear envelope. To date, the mechanism of the association of human cytomegalovirus (HCMV) capsids with the NEC, which in turn initiates the specific steps of nuclear capsid budding, remains undefined. Here, we provide electron microscopy-based data demonstrating the association of both nuclear capsids and NEC proteins at nuclear lamina budding sites. Specifically, immunogold labelling of the core NEC constituent pUL53 and NEC-associated viral kinase pUL97 suggested an intranuclear NEC-capsid interaction. Staining patterns with phospho-specific lamin A/C ...

Abstract: The nuclear phase of herpesvirus replication is regulated through the formation of regulatory multi-component protein complexes. Viral genomic replication is followed by nuclear capsid assembly, DNA encapsidation and nuclear egress. The latter has been studied intensely pointing to the formation of a viral core nuclear egress complex (NEC) that recruits a multimeric assembly of viral and cellular factors for the reorganization of the nuclear envelope. To date, the mechanism of the association of human cytomegalovirus (HCMV) capsids with the NEC, which in turn initiates the specific steps of nuclear capsid budding, remains undefined. Here, we provide electron microscopy-based data demonstrating the association of both nuclear capsids and NEC proteins at nuclear lamina budding sites. Specifically, immunogold labelling of the core NEC constituent pUL53 and NEC-associated viral kinase pUL97 suggested an intranuclear NEC-capsid interaction. Staining patterns with phospho-specific lamin A/C ...

The nuclear phase of herpesvirus replication is regulated through the formation of regulatory multi-component protein complexes. Viral genomic replication is followed by nuclear capsid assembly, DNA encapsidation and nuclear egress. The latter has been studied intensely pointing to the formation of a viral core nuclear egress complex (NEC) that recruits a multimeric assembly of viral and cellular factors for the reorganization of the nuclear envelope. To date, the mechanism of the association of human cytomegalovirus (HCMV) capsids with the NEC, which in turn initiates the specific steps of nuclear capsid budding, remains undefined. Here, we provide electron microscopy-based data demonstrating the association of both nuclear capsids and NEC proteins at nuclear lamina budding sites. Specifically, immunogold labelling of the core NEC constituent pUL53 and NEC-associated viral kinase pUL97 suggested an intranuclear NEC-capsid interaction. Staining patterns with phospho-specific lamin A/C antibodies are

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TY - JOUR. T1 - Derivation of a triple mosaic adenovirus based on modification of the minor capsid protein IX. AU - Tang, Yizhe. AU - Le, Long P.. AU - Matthews, Qiana L.. AU - Han, Tie. AU - Wu, Hongju. AU - Curiel, David T.. N1 - Funding Information: Ad5IXFlag and Ad5IXHis 6 are kind gifts from Dr. Anton Borovjagin, the stable cell line expressing sCAR protein is a kind gift from Dr. Maaike Everts. Discussions with Dr. Hideyo Ugai, Dr. Anton Borovjagin and Dr. Igor Dmitriev were helpful and stimulating throughout this work. The authors also thank Dr. Lacey McNally for technical support of immunogold electron microscopy and Minghui Wang for assistance in quantitative PCR analysis. This work was supported by the following grants: NIH R01CA111569 (Dr. David T. Curiel), NIH 5T32AI07493-11 (Dr. Casey Morrow), Juvenile Diabetes Research Foundation 1-2005-71 (Dr. Hongju Wu) and 5-2007-660 (Dr. Hongju Wu). PY - 2008/8/1. Y1 - 2008/8/1. N2 - Adenoviral capsid protein IX (pIX) has been shown to be a ...

The mature HIV-1 conical core formation proceeds through highly regulated protease cleavage of the Gag precursor, which ultimately leads to substantial rearrangements of the capsid (CAp24) molecule involving both inter- and intra-molecular contacts of the CAp24 molecules. In this aspect, Asp51 which is located in the N-terminal domain of HIV-1 CAp24 plays an important role by forming a salt-bridge with the free imino terminus Pro1 following proteolytic cleavage and liberation of the CAp24 protein from the Pr55Gag precursor. Thus, previous substitution mutation of Asp51 to alanine (D51A) has shown to be lethal and that this invariable residue was found essential for tube formation in vitro, virus replication and virus capsid formation. We extended the above investigation by introducing three different D51 substitution mutations (D51N, D51E, and D51Q) into both prokaryotic and eukaryotic expression systems and studied their effects on in vitro capsid assembly and virus infectivity. Two substitution

Recombinant adeno-associated viruses (AAVs) are quickly becoming the preferred viral vector for viral gene delivery for the treatment of a wide variety of genetic disorders. However, since their use in a clinical trial targeting hemophilia B patients 10 years ago, immune responses to the AAV capsid appear to have hampered some of the early clinical gene transfer efficacy. Indeed, AAV-based gene transfer has been shown to reactivate capsid-specific memory T cells, which have correlated with a decline in AAV-transduced tissue in some patients. Importantly, clinical trials have also shown that this reactivation can be quelled by administering time-course taper of glucocorticoid steroids before or after dosing. More recently, two clinical studies have shown that AAV gene transfer is not only able to induce a deleterious immune response, but also can result in the initiation of a tolerance to the AAV capsid mediated by regulatory T cells and exhausted T cells. This article reviews clinical trials describing

Peptides from different regions of the poliovirus type 1 capsid protein VP1 were synthesized. Antibodies raised against these peptides in rabbits and rats recognized the cognate peptides and denatured VP1. Peptides from four regions of VP1 generated antisera with neutralizing titers specifically against poliovirus type 1. Antisera against all other regions of VP1 failed to neutralize virus infectivity, although some of the antisera clearly bound to native virions. Thus, the neutralizing determinants on VP1 reside in specific noncontiguous regions of the protein and can be defined by specific peptides from these regions. ...

Viral particles derived from mouse polyomavirus can be potentially used as a delivery system for therapeutic genes and drugs into target cells. This thesis focuses on preparation and characterization of polyomaviral particles that are modified with cell-penetrating peptides in order to increase efficiency of transduction of reporter genes into human cells. Viral particles that are composed of major capsid protein VP1 in combination with minor capsid protein VP2 and minor capsid protein VP3 that is modified with octaarginine, LAH4 peptide or with transduction domain of adenoviral protein VI are analysed in transduction assays. The thesis also provides information about the effect of the modification on encapsidation of heterologous DNA. The results of transduction assays performed with modified particles containing encapsidated luciferase gene revealed that efficiency of transduction did not increase but decreased in comparison with unmodified particles. These findings help to elucidate the role ...

Author Summary Cellular and viral life cycles are connected through multiple, though poorly understood, mechanisms. Parvoviruses infect humans and a broad spectrum of animals, causing a variety of diseases, but they are also used in experimental cancer therapy and serve as vectors for gene therapy. Parvoviruses can only multiply in proliferating cells providing essential replicative and transcriptional functions. However, it is unknown whether the cell cycle regulatory machinery may also control parvovirus assembly. We found that the nuclear translocation of parvovirus MVM capsid subunits (VPs) was highly dependent on physiological cell cycle regulations in mammalian fibroblasts, including: quiescence, progression through G1/S boundary, DNA synthesis, and cell to cell contacts. VPs nuclear translocation was significantly more sensitive to cell cycle controls than viral genome replication and gene expression. The results support nuclear capsid assembly as the major driving process of parvoviruses

John Walters work encompasses a diverse range of media, including painting, drawing, artists books, sculpture, costume, performance, video, sound, installation and spatial design. His oeuvre is characterised by an exuberant use of colour and pattern as well as an absurdist and tragicomic use of humour. He works serially, producing iterative bodies of work that accumulate to form large and distinct projects, and often collaborates with individuals and institutions such as other artists, scientists and museums in order to exchange images, ideas and narratives. CAPSID (2018-19) incorporated 250 artworks, and was the result of a collaboration between the artist and molecular virologist Professor Greg Towers of University College London. The exhibition addressed a crisis of representation surrounding viruses such as HIV, by bringing new scientific knowledge about viral capsids to the attention of the wider public. Capsids are protein shells contained within viruses that help protect and deliver ...

John Walters work encompasses a diverse range of media, including painting, drawing, artists books, sculpture, costume, performance, video, sound, installation and spatial design. His oeuvre is characterised by an exuberant use of colour and pattern as well as an absurdist and tragicomic use of humour. He works serially, producing iterative bodies of work that accumulate to form large and distinct projects, and often collaborates with individuals and institutions such as other artists, scientists and museums in order to exchange images, ideas and narratives. CAPSID (2018-19) incorporated 250 artworks, and was the result of a collaboration between the artist and molecular virologist Professor Greg Towers of University College London. The exhibition addressed a crisis of representation surrounding viruses such as HIV, by bringing new scientific knowledge about viral capsids to the attention of the wider public. Capsids are protein shells contained within viruses that help protect and deliver ...

The levels of c-myc, c-fos, and JE mRNAs accumulate in a biphasic pattern following infection of quiescent BALB/c 3T3 mouse cells with polyomavirus. Maximal levels of c-myc and c-fos mRNAs were seen within 1 hr and were nearly undetectable at 6 hr after infection. At 12 hr after infection mRNA levels were again maximal and remained elevated thereafter. Empty virions (capsids) and recombinant VP1 protein, purified from Escherichia coli, induced the early but not the late phase of mRNA accumulation. Virions, capsids, and recombinant VP1 protein stimulated [3H]thymidine nuclear labeling and c-myc mRNA accumulation ina dose-responsive manner paralleling their affinity for the cell receptor for polyoma. The second phase of mRNA accumulation is regulated by the viral early gene products, as shown by polyomavirus early gene mutants and by a transfected cell line (336a) expressing middle tumor antigen upon glucocorticoid addition. These results suggest that polyomavirus interacts with the cell membrane ...

Self-assembly is a vital part of the of the RNA virus life cycle. The assembly of viral coat proteins around viral RNA occurs both in vivo and in vitro, suggesting that viral capsid assembly may be driven by a minimization of free energy. To better understand this process, we modify the interactions between coat proteins and between the coat proteins and RNA of MS2 bacteriophage in vitro by varying the ionic strength and pH, and we study the assembly using dynamic and static light scattering. From dynamic light scattering we determine the assembly yield and the size distribution of assembled products. From static light scattering, we measure the kinetics of assembly in bulk. By comparing the results from these two different techniques to each other and to results from gel electrophoresis, we infer features of the assembly pathway. ...

Author: Walz, J. et al.; Genre: Journal Article; Published in Print: 1999; Keywords: Electron microscopy; Icosahedral capsids; Image analysis; Thermoplasma.; Degradation; Microscopy; Resolution; Products.; Biochemistry & Biophysics in Current Contents(R)/Life Sciences.; Title: Capsids of tricorn protease studied by electron cryomicroscopy

Background: Human papillomavirus (HPV)-45 genotype circulates in high percentage in Bandung area - Indonesia, after HPV-16 and HPV-18. The aim of this study was to analyse variations of major capsid (L1) HPV-45 and its phylogeny. Furthermore in silico protein structure and epitope prediction was explored. Methods: L1 gene of HPV-45 was amplified, sequenced and aligned. Phylogenetic tree had been built and compared with a complete L1 HPV-45 sequence. Structure and epitope prediction of L1 protein were then developed in silico. Results: Of 5 L1 HPV-45 sequences collected, we have detected one variant of sub lineage A2 which was considered as a new variant, and two variants of B2. Superimposition of structure of these two variants with reference showed very similar structure. Furthermore, seven amino acid substitutions were found within these L1 variants of which two substitutions might change the polarity of corresponding amino acid I329T and S383G. The S383G occurred in surface loop (HI-Loop) of new L1

3D cryo-EM image of Rice Dwarf Virus (RDV) processed via our meshing pipeline. (a): two dimensional projection of an acquired cryo-EM image of the virus particles. (b): reconstructed 3D image of intensities -bisected to show interior and volume rendered. (c): the RDV capsid with inner core packing of proteins and nucleic acids, volume rendered with appropriate opacity transfer function. (d): RDV shown in (c) after contrast enhancement step. (e): segmentation before filtering into an outer capsid layer (blue), inner capsid layer (purple) and the inner core (green). Note noise around the outer capsid. (f): segmentation applied after anisotropic filtering of the 3D image. (g): outer capsid layer is further automatically segmented into locally asymmetric subunits. A P8 trimeric subunit is identified. (h): surface rendering of the segmented P8 trimeric subunit from the outer capsid. (i): the P8 trimeric subunits is further segmented into three P8 monomeric proteins (colored differently). Triangular ...

Fingerprint Dive into the research topics of Mature HIV-1 capsid structure by cryo-electron microscopy and all-atom molecular dynamics. Together they form a unique fingerprint. ...

Cloning and expression of the gene encoding for the major capsid protein, VP-1, of the D-variant of encephalomyocarditis virus in Escherichia ...

Purified Recombinant Rubella Virus Capsid (C) Protein from Creative Biomart. Recombinant Rubella Virus Capsid (C) Protein can be used for research.

PepTivator® Zika Capsid Protein C is a pool of lyophilized peptides, consisting mainly of 15-mer sequences with 11 amino acids overlap, covering the sequence of the Zika capsid protein C domain from virus isolate Z1106033 polyprotein gene (GenBank Acc. no. KU312312.1).In vitro stimulation of antigen-specific T cells with PepTivator Peptide Pools causes the secretion of effector cytokines and the up-regulation of activation markers, which then allow the detection and isolation of antigen-specific T cells. | Singapore

The group of CAO Yu used the capsid protein (HaCP) of Baculovirus expressed by E. coli and found by cryo-electron microscopy that a variety of exogenous proteins expressed at the N-terminus of HaCP could be directly fused and displayed on the outer surface. HaCP had two forms of assembly in vitro: one with a small diameter named N-tube, and another with a relatively large diameter, named W-tube. The N-tube is a helical structure similar to the TMV capsid, with 111/3 protein subunits arranged in a spiral along each circle. The W-tube is a stack of disc structures consisting of 12 protein subunits with a 11.25° rotation between adjacent discs. The interaction between the HaCP nanotube subunits was tunable, leading to high flexibility of the resulting nanotubes. HaCP nanotubes thus allow to accommodate larger guest compounds compared to assembled subunits of TMVs and thus might be evolved into nanoscale functional molecules.. CAS news release, July 30, 2018. ...

Guest editorial: Technology to meet the challenge of efficiency in the regulated AAV vector space. In this guest editorial, based on a blog post by Gyros Protein Technologies, learn how scientists at AstraZeneca developed a reliable method for adeno-associated virus (AAV) capsid titer quantification.. Vectors based on AAV are used widely in gene therapy for in vivo gene delivery. The success of AAV-based therapeutic production depends on the availability of laboratory tools that can efficiently deliver reliable data, including the determination of virus particle titer (physical titer). To achieve this, a team at AstraZeneca searched for an immunoassay platform that could improve on ELISA in terms of dynamic range, sample volume, and productivity. Its evaluation showed that Gyrolab systemmet its needs for AAV process development, with a more reliable measurement of AAV capsid titer for in-process and purified samples.. Gene therapy has seen a massive explosion in activity since the first clinical ...

Fingerprint Dive into the research topics of A spectrum of HRT-dependent hypersensitive responses elicited by the 52 amino acid N-terminus of turnip crinkle virus capsid protein and its mutants. Together they form a unique fingerprint. ...

The structures of Penicillium stoloniferum viruses F and S (PsV-F and PsV-S) have been determined by X-ray crystallography (PsV-F) and electron cryomicroscopy (PsV-F and PsV-S) (Ochoa et al., 2008, Pan et al., 2009). The outer diameter of the capsid is about 35-40 nm. The most prominent features of the capsid are 60 arch-like protrusions that decorate a spherical shell (Figure 1.Gammapartitivirus.A-C). Each particle contains 120 CP molecules arranged with icosahedral symmetry. The tertiary structure of each CP molecule consists of two distinct domains, one of which forms the continuous, 3 nm thick capsid, and the other that, with the corresponding domain of a neighboring CP, comprises the arch (Figure 1.Gammapartitivirus.E-F). The asymmetric unit of the icosahedron consists of two related CP molecules, CPA and CPB. The 60 CPA molecules are organized as flower-like pentamers, each centered about one of the 12 vertices of the icosahedral capsid (Figure 1.Gammapartitivirus.A). The 60 CPB molecules ...

Antigen Background The Caliciviridae are a family of positive-stranded RNA viruses of unique morphology characterized by a series of cup-like depressions on the surface of the virus. Feline Calicivirus (FCV) is a ubiquitous pathogen of cats producing a variety of clinical symptoms, including oral ulceration, upper respiratory tract infection and polyarthritis. FCV has a genome of 7.7kb which encodes several proteins. Product Specific Information NCL-1G9 detects one of these, a capsid protein of 62 kD.. ...

TY - JOUR. T1 - NONO Detects the Nuclear HIV Capsid to Promote cGAS-Mediated Innate Immune Activation. AU - Lahaye, Xavier. AU - Gentili, Matteo. AU - Silvin, Aymeric. AU - Conrad, Cécile. AU - Picard, Léa. AU - Jouve, Mabel. AU - Zueva, Elina. AU - Maurin, Mathieu. AU - Nadalin, Francesca. AU - Knott, Gavin J.. AU - Zhao, Baoyu. AU - Du, Fenglei. AU - Rio, Marlène. AU - Amiel, Jeanne. AU - Fox, Archa H.. AU - Li, Pingwei. AU - Etienne, Lucie. AU - Bond, Charles S.. AU - Colleaux, Laurence. AU - Manel, Nicolas. PY - 2018/10/4. Y1 - 2018/10/4. N2 - Detection of viruses by innate immune sensors induces protective antiviral immunity. The viral DNA sensor cyclic GMP-AMP synthase (cGAS) is necessary for detection of HIV by human dendritic cells and macrophages. However, synthesis of HIV DNA during infection is not sufficient for immune activation. The capsid protein, which associates with viral DNA, has a pivotal role in enabling cGAS-mediated immune activation. We now find that NONO is an ...

To characterize the immunity developed by patients infected by chikungunya virus (CHIKV), we studied the intensity and specificity of CHIKV-specific T cells mediated responses in chronic and recovered patients at 12 to 24 months post-infection. T cells were challenged in vitro against CHIKV synthetic peptides covering the length of three viral proteins, capsid, E2 and nsP1 proteins as well as all inactivated virus particles. Cytokine production was assessed by ELISPOT and intracellular labeling. T cells producing IFN-γ were detected against CHIKV in 85% patients cells either by direct ELISPOT assay (69% of patients) or after expansion of memory T cells allowing the detection of both CD4 and CD8 specific-T cells in 16% additional cases. The IFN-γ response was mainly engaged in response to nsP1 or E2 (52% and 46% cases, respectively) but in only 27% cases against the capsid. The anti-E2 response represented half the magnitude of the total CHIKV IFN-γ production and was mainly directed against the C

Abstract. Thermal shift assays measure the stability of macromolecules and macromolecular assemblies as a function of temperature. The Particle Stability Thermal Release Assay (PaSTRy) of picornaviruses is based on probes becoming strongly fluorescent upon binding to hydrophobic patches of the protein capsid (e.g., SYPRO Orange) or to the viral RNA genome (e.g., SYTO-82) that become exposed upon heating virus particles. PaSTRy has been exploited for studying the stability of viral mutants, viral uncoating, and the effect of capsid-stabilizing compounds. While the results were usually robust, the thermal shift assay with SYPRO Orange is sensitive to surfactants and EDTA and failed at least to correctly report the effect of excipients on an inactivated poliovirus 3 vaccine. Furthermore, interactions between the probe and capsid-binding antivirals as well as mutual competition for binding sites cannot be excluded. To overcome these caveats, we assessed differential scanning fluorimetry with a ...

An alliance has been formed between SIRION Biotech and Heidelberg University Hospital for the development of novel AAV capsids and technologies addressing shortcomings of clinical gene therapy trials.

These results indicate that the mouse Fv1 gene has had antiviral activity since shortly after its introduction into the Mus genome, an event that occurred during a period of rapid diversification that gave rise to the 4 Mus subgenera ≈7 MY ago (10, 23). That this sequence has long had antiviral function is supported by the observation that the Fv1 genes of 2 species of Nannomys have novel Fv1-mediated antiviral phenotypes, and by the demonstration that Fv1 has been under positive Darwinian selection. We identified 2 segments of Fv1 potentially involved in restriction: an MHR segment known to function in capsid binding, and 3 residues in the C-terminal domain (CTD) of which 2 are known to contribute to the specificity of Fv1 restriction in laboratory mice.. The mechanism of Fv1 restriction has not been elucidated after 40 years of investigation, but the determination that Fv1 encodes a retroviral capsid-like protein suggests that Fv1 may bind capsids of exogenous virus and interfere with capsid ...

Crystal structure of a distinct viral capsid/ssDNA complex illuminates how viruses assemble into infectious virions Conference Paper ...

An interaction between the HSV-1 UL25 capsid protein and cellular microtubule-associated protein was found using a yeast two-hybrid screen and β-D-galactosidase activity assays. Immunofluorescence microscopy of the UL25 protein demonstrated its co-localization with cellular microtubule-associated protein in the plasma membrane. Further investigations with deletion mutants suggest that UL25 is likely to have a function in the nucleus.

Alpha herpesviruses, such as herpes simplex virus and pseudorabies virus (PRV), are neuroinvasive dsDNA viruses that establish life-long latency in peripheral nervous system (PNS) neurons of their native hosts. Following reactivation, the infection can spread back to the initial mucosal site of infection or, in rare cases, to the central nervous system with usually serious outcomes. During entry and egress, viral capsids depend on microtubule-based molecular motors for efficient and fast transport. In axons of PNS neurons, cytoplasmic dynein provides force for retrograde movements towards the soma, and kinesins move cargo in the opposite, anterograde direction. The dynamic properties of virus particles in cells can be imaged by fluorescent protein fusions to the small capsid protein VP26, which are incorporated into capsids. However, single-color fluorescent protein tags fail to distinguish virus inoculum from progeny. Therefore, we established a dual-color system by growing a recombinant PRV ...

The host protein cyclophilin A (CypA) can both stimulate and inhibit HIV-1 infection through its interaction with the viral capsid (CA). CypA enhances the early stages of HIV-1 infection in part by promoting nuclear import of the virus; while the details of its ability to inhibit HIV-1 infection are less clear. This thesis advances our understanding of the mechanisms underlying the ability of CypA to inhibit HIV-1 infection. I demonstrate that CypA inhibits nuclear import of HIV-1 in the presence of inhibitory capsid-binding host proteins TRIM5α and CPSF6, and that inhibition is not a consequence of increased binding of inhibitory factors to the viral capsid. My work also demonstrates that CypA-dependent inhibition depends in part, on a conserved domain of HIV-1 CA which determines interactions with host nuclear pore proteins. These results suggest a common mechanism underlies the ability of CypA to stimulate HIV-1 infectivity in some cells and to inhibit infection in others. ...

DNA may scrunch like a worm to get inside viral shells, a team including Georgia Tech researchers reports in the Journal of Physical Chemistry B. This deeper understanding could lead to new ways to fight pathogens and design powerful DNA transporters.. A critical step in viral replication is the packaging of genetic material. To successfully invade host cells, viral particles must hijack the hosts machinery to make copies of viral genetic material and build protein shells called capsids to house viral DNA or RNA. Scientists have been studying how the genetic material is driven into capsids so they might one day block this step.. Viral capsids are assembled from a number of identical protein subunits, like a soccer ball sewn together from panels. At the lone opening sits a protein complex, called the protein tunnel, through which DNA enters and exits the capsid, analogous to the air valve that allows a soccer ball to be inflated.. The proteins driving this process are among the strongest ...

TY - JOUR. T1 - Genetic diversity in the major capsid L1 protein of HPV-16 and HPV-18 in the Netherlands. AU - Medical Microbiological Laboratories and Municipal Health Services. AU - King, Audrey J. AU - Sonsma, Jan A. AU - Vriend, Henrike J. AU - van der Sande, Marianne A B. AU - Feltkamp, Mariet C. AU - Boot, Hein J. AU - Koopmans, Marion P G. N1 - CPDF. PY - 2016. Y1 - 2016. N2 - OBJECTIVES: Intratypic molecular variants of human papillomavirus (HPV) type-16 and -18 exist. In the Netherlands, a bivalent vaccine, composed of recombinant L1 proteins from HPV-16 and -18, is used to prevent cervical cancer since 2009. Long-term vaccination could lead to changes in HPV-16 and -18 virus population, thereby hampering vaccination strategies. We determined the genetic diversity of the L1 gene in HPV-16 and -18 viral strains circulating in the Netherlands at the start of vaccination in order to understand the baseline genetic diversity in the Dutch population.METHODS: DNA sequences of the L1 gene were ...

Icosahedral symmetry is equivalently the projective special linear group PSL(2,5), and is the symmetry group of the modular curve X(5), and more generally PSL(2,p) is the symmetry group of the modular curve X(p). The modular curve X(5) is geometrically a dodecahedron with a cusp at the center of each polygonal face, which demonstrates the symmetry group. This geometry, and associated symmetry group, was studied by Felix Klein as the monodromy groups of a Belyi surface - a Riemann surface with a holomorphic map to the Riemann sphere, ramified only at 0, 1, and infinity (a Belyi function) - the cusps are the points lying over infinity, while the vertices and the centers of each edge lie over 0 and 1; the degree of the covering (number of sheets) equals 5. This arose from his efforts to give a geometric setting for why icosahedral symmetry arose in the solution of the quintic equation, with the theory given in the famous (Klein 1888); a modern exposition is given in (Tóth 2002, Section 1.6, ...