Gag-Pol polyprotein: Mediates, with Gag polyrotein, the essential events in virion assembly, including binding the plasma membrane, making the protein-protein interactions necessary to create spherical particles, recruiting the viral Env proteins, and packaging the genomic RNA via direct interactions with the RNA packaging sequence (Psi). Gag-Pol polyprotein may regulate its own translation, by the binding genomic RNA in the 5-UTR. At low concentration, the polyprotein would promote translation, whereas at high concentration, the polyprotein would encapsidate genomic RNA and then shutt off translation.

Gag-Pol polyprotein and Gag polyprotein may regulate their own translation, by the binding genomic RNA in the 5-UTR. At low concentration, Gag-Pol and Gag would promote translation, whereas at high concentration, the polyproteins encapsidate genomic RNA and then shutt off translation (By similarity).

Foamy viruses (FVs) differ from all other genera of retroviruses (orthoretroviruses) in many aspects of viral replication. In this review, we discuss FV assembly, with special emphasis on Pol incorporation. FV assembly takes place intracellularly, near the pericentriolar region, at a site similar to that used by betaretroviruses. The regions of Gag, Pol and genomic RNA required for viral assembly are described. In contrast to orthoretroviral Pol, which is synthesized as a Gag-Pol fusion protein and packaged through Gag-Gag interactions, FV Pol is synthesized from a spliced mRNA lacking all Gag sequences. Thus, encapsidation of FV Pol requires a different mechanism. We detail how WT Pol lacking Gag sequences is incorporated into virus particles. In addition, a mutant in which Pol is expressed as an orthoretroviral-like Gag-Pol fusion protein is discussed. We also discuss temporal regulation of the protease, reverse transcriptase and integrase activities of WT FV Pol.

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Retroviral proteins coded by the pol gene. They are usually synthesized as a protein precursor (POLYPROTEINS) and later cleaved into final products that include reverse transcriptase, endonuclease/integrase, and viral protease. Sometimes they are synthesized as a gag-pol fusion protein (FUSION PROTEINS, GAG-POL). pol is short for polymerase, the enzyme class of reverse transcriptase. . ...

The polypeptide composition of HIV-I virus-like particles produced by CV-I cells during mono- and coinfection with recombinant vaccinia virus (rVV) strains containing the whole (p55) and carboxyterminal truncated (p48) gag genes and gag-pol sequence is studied. In monoinfection both the gag-strains actively produced virus-like particles consisting of non-processed p55Gag and p48Gag polyprotein without p6 domain. In case of a coinfection of the cells with one of these strains and the rVV producing p160Gag-Pol polyprotein the virus-like particles consisted of p24 protein and a negligible amount of non-processed Gag precursors. The share of p24 protein increased in proportion to the duration of coinfection and decreased with a reduction of multiplicity of infection with rVV carrying p160Gag-Pol. Hence, the absence of p6 domain does not influence the processing of Gag proteins during virus-like particles assembly and budding. In contrast to the natural systems of HIV-I development, in the rVV expression

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Within the family of Retroviridae, foamy viruses (FVs) are unique and unconventional with respect to many aspects in their molecular biology, including assembly and release of enveloped viral particles. Both components of the minimal assembly and release machinery, Gag and Env, display significant differences in their molecular structures and functions compared to the other retroviruses. This led to the placement of FVs into a separate subfamily, the Spumaretrovirinae. Here, we describe the molecular differences in FV Gag and Env, as well as Pol, which is translated as a separate protein and not in an orthoretroviral manner as a Gag-Pol fusion protein. This feature further complicates FV assembly since a specialized Pol encapsidation strategy via a tripartite Gag-genome–Pol complex is used. We try to relate the different features and specific interaction patterns of the FV Gag, Pol, and Env proteins in order to develop a comprehensive and dynamic picture of particle assembly and release, but also

POL_HV1B1] Gag-Pol polyprotein and Gag polyprotein may regulate their own translation, by the binding genomic RNA in the 5-UTR. At low concentration, Gag-Pol and Gag would promote translation, whereas at high concentration, the polyproteins encapsidate genomic RNA and then shutt off translation (By similarity).[1] Matrix protein p17 has two main functions: in infected cell, it targets Gag and Gag-pol polyproteins to the plasma membrane via a multipartite membrane-binding signal, that includes its myristoylated N-terminus. The second function is to play a role in nuclear localization of the viral genome at the very start of cell infection. Matrix protein is the part of the pre-integration complex. It binds in the cytoplasm the human BAF protein which prevent autointegration of the viral genome, and might be included in virions at the ration of zero to 3 BAF dimer per virion. The myristoylation signal and the NLS thus exert conflicting influences its subcellular localization. The key regulation ...

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The PDB archive contains information about experimentally-determined structures of proteins, nucleic acids, and complex assemblies. As a member of the wwPDB, the RCSB PDB curates and annotates PDB data according to agreed upon standards. The RCSB PDB also provides a variety of tools and resources. Users can perform simple and advanced searches based on annotations relating to sequence, structure and function. These molecules are visualized, downloaded, and analyzed by users who range from students to specialized scientists.

The PDB archive contains information about experimentally-determined structures of proteins, nucleic acids, and complex assemblies. As a member of the wwPDB, the RCSB PDB curates and annotates PDB data according to agreed upon standards. The RCSB PDB also provides a variety of tools and resources. Users can perform simple and advanced searches based on annotations relating to sequence, structure and function. These molecules are visualized, downloaded, and analyzed by users who range from students to specialized scientists.

Translation of MuLV and MSV RNAs in nuclease-treated reticulocyte extracts: enhancement of the gag-pol polypeptide with yeast suppressor tRNA ...

This tool takes as input a set of Gag-Pol sequences including the gene for the viral PR and one or more of its CSs, and produces a prediction for potential secondary drug resistance mutations in the latter. The cleavage of the Gag-Pol polyprotein by the PR is essential for the infectivity of HIV virions. PI therapy can give rise to primary resistance mutations in the PR, which are often associated with a decreased activity of the enzyme. Impaired function can be partially restored by compensatory mutations in the CS, probably by providing a better substrate for the mutated proteases. Associated pairs of mutations have been detected from large sets of sequences (populations of molecules) by covariation analysis (Hoffman et al. 2003), and we have extended this approach to identify CS mutations that arise in response to primary resistance mutations in the PR. We analysed HIV-1 subtype B nucleotide sequences containing the protease region from the Los Alamos HIV Sequence Database ...

TY - JOUR. T1 - Recombinant tagging system using ribosomal frameshifting to monitor protein expression. AU - Han, Se Jong. AU - Cho, Sayeon. AU - Lowehhaupt, Ky. AU - Park, So Young. AU - Sim, Sang Jun. AU - Kim, Yang Gyun. PY - 2013/3/1. Y1 - 2013/3/1. N2 - For rapid and accurate quantitation of recombinant proteins during expression and after purification, we introduce a new tagging strategy that expresses both target proteins and limitedly tagged target proteins together in a single cell at a constant ratio by utilizing cis-elements of programmed -1 ribosomal frameshifting (-1RFS) as an embedded device. -1RFS is an alternative reading mechanism that effectively controls protein expression by many viruses. When a target gene is fused to the enhanced green fluorescent protein (EGFP) gene with a -1RFS element implanted between them, the unfused target and the target-GFP fusion proteins are expressed at a fixed ratio. The expression ratio between these two protein products is adjustable simply by ...

An intact PR domain is known to be required for RIZ1 tumor suppression function in mice (18) . RIZ1 knockout mice are tumor prone and are deficient only in the PR domain as these animals express the PR-deficient product RIZ2. The presence of naturally occurring mutations in the PR domain that altered RIZ1 transcription factor function additionally suggests an important role for this domain in human cancer (18) . These observations, taken together with the present findings of HMT activity for the PR domain and the inactivation of this activity by naturally occurring mutations, suggest that HMT activity is important to RIZ1 tumor suppression function. The H3-K9 methylation activity of RIZ1 may play a similar role as other HMTs in the epigenetic control of gene silencing. Several HMTs are known to be key players in the mitotic inheritance of cell fates and gene expression patterns (33, 34, 35, 36, 37, 38) . Some of these enzymes are commonly involved in cancer (10) . Altered epigenetic control of ...

HEK 293T/17 cells were transformed with adenovirus E1a carrying a temperature sensitive T antigen co-selected with neomycin. Transformation was brought about by the insertion of approximately 4.5 kilobases of viral genome into human chromosome 19. Gag-pol was introduced with hygromycin as the co-selectable marker and the envelope proteins were introduced with diptheria resistance as the co-selectable marker.

HEK 293T/17 cells were transformed with adenovirus E1a carrying a temperature sensitive T antigen co-selected with neomycin. Transformation was brought about by the insertion of approximately 4.5 kilobases of viral genome into human chromosome 19. Gag-pol was introduced with hygromycin as the co-selectable marker and the envelope proteins were introduced with diptheria resistance as the co-selectable marker.

TY - JOUR. T1 - The Complete Sequence of Leishmania RNA Virus LRV2-1, a Virus of an Old World Parasite Strain. AU - Scheffter, Scott M.. AU - Ro, Young T.. AU - Chung, In K.. AU - Patterson, J. L.. PY - 1995/9/10. Y1 - 1995/9/10. N2 - A complete cDNA sequence is reported for LRV2-1, the first Leishmania RNA virus known to infect an Old World parasite, Leishmania major. Sequence analyses show that LRV2-1 differs significantly from members of the LRV1 genus which infect New World parasites. The data support a view that transmission of LRV is strictly vertical and suggest that LRV predate the divergence of Old and New World parasites. As a consequence of this divergence, conserved features can be identified for the first time in Leishmania virus proteins. A finding that the virus capsid and polymerase genes do not overlap is unique among the known Totiviridae and infers that a gag-pol fusion protein cannot be produced simply via tRNA slippage in LRV2-1.. AB - A complete cDNA sequence is reported ...

Vectors derived from human immunodeficiency virus type 1 (HIV-1) appear an attractive option for many gene therapy applications. This is due to their ability to transduce noncycling cell populations and to integrate their genome into the host cell chromosome, resulting in the stable genetic modification of the transduced cell. These properties have permitted the direct in vivo transduction of several tissues, including the central nervous system, retina, and liver. However, the pathogenic nature of HIV-1 has raised considerable concerns about the safety of such vector systems. To help address these concerns, we have expressed each of the primary transcriptional units encoding trans functions relevant for vector production in individual plasmid constructs. The gag-pol gene sequence was codon-optimized for expression in mammalian cells resulting in high level Rev/Rev-response element (RRE)-independent expression. Codon optimization of gag-pol also reduces sequence homology with vectors containing ...

1. Esnault C, Heidmann O, Delebecque F, Dewannieux M, Ribet D, Hance AJ, et al. APOBEC3G cytidine deaminase inhibits retrotransposition of endogenous retroviruses. Nature. 2005;433(7024):430-3. Epub 2005/01/28. doi: 10.1038/nature03238 15674295.. 2. Harris RS, Bishop KN, Sheehy AM, Craig HM, Petersen-Mahrt SK, Watt IN, et al. DNA deamination mediates innate immunity to retroviral infection. Cell. 2003;113(6):803-9. Epub 2003/06/18. doi: 10.1016/s0092-8674(03)00423-9 12809610.. 3. Mangeat B, Turelli P, Caron G, Friedli M, Perrin L, Trono D. Broad antiretroviral defence by human APOBEC3G through lethal editing of nascent reverse transcripts. Nature. 2003;424(6944):99-103. Epub 2003/06/17. doi: 10.1038/nature01709 12808466.. 4. Suspene R, Aynaud MM, Koch S, Pasdeloup D, Labetoulle M, Gaertner B, et al. Genetic editing of herpes simplex virus 1 and Epstein-Barr herpesvirus genomes by human APOBEC3 cytidine deaminases in culture and in vivo. J Virol. 2011;85(15):7594-602. Epub 2011/06/03. doi: ...

If a sequence of interest is not currently in the PRFdb, it can be imported via its NCBI accession number. Sequences added in this manner will be filtered within hours of import. Sequences imported into the PRFdb are also folded using sequential windows across the entire sequence in order to create a graphical minimum free energy landscape. This enables users to submit longer or shorter sequence strings for computational folding, a particularly useful feature e.g. for eliminating extraneous sequence that may not be involved in actual RNA folding. For example, the computational analysis of the 100 nucleotide sequence downstream of the slippery site of the mouse Ma3 -1 PRF signal provided by the PRFdb predicts tandem stem loop structures. However, when only 55 nt of downstream sequence are provided, PRFdB predicts the empirically documented pseudoknot structure [14].. Sequences to be analyzed by the PRFdb are imported into the database, filtered using RNAMotif [15], folded with secondary ...

Analysis of frameshifting in the viral context. (A) Radiolabeled TMEV translation products. BHK-21 cells were infected with either WT, SCM, SS, or LVWT viruses

CFG1 is a Ty3/Gypsy LTR retrotransposon described in the genome of the mollusca Chlamys Farreri ( Wang et al 2007). Expression of CFG1 appears to be controlled by a post-transcriptional gene silencing mechanisms associated to reverse transcriptase (RT) methylation ( Wang et al 2007). CFG1 belongs, along with other LTR retrotransposons, to Mag a large Ty3/Gypsy cluster widely distributed in both protostomes and deuterostomes (for more details, see Malik and Eickbush 1999; Volff et al. 2001; Tubio, Naveira and Costas 2004). The genomic structure of CFG1 is 4.7 Kb in size including LTRs of 192 and 189 nt. The internal region of this element displays a Primer Binding Site (PBS), a single Open Reading Frame (ORF) that codes for a single gag-pol polyprotein, and a Polypurine Tract (PPT) adjacent to the 3´LTR ( Wang et al 2007). ...

Summary Retroviral packaging cell lines were constructed by using the gag-pol gene of spleen necrosis virus, the gag-pol gene of Moloney murine leukaemia virus and the env gene of bovine leukaemia virus. The plasmids containing the gag-pol genes and the plasmid containing the env gene were cotransfected into NIH/3T3 and D17 cells. The cells containing the helper virus constructs were tested for their ability to package replication-defective murine leukaemia and avian reticuloendotheliosis retrovirus vectors. The titre of vector virus produced by each of the retroviral packaging cell lines was about 102 colony-forming units per ml of medium. Tests for events that might result in intact replication-competent retroviruses showed no evidence for the generation of such viruses. The vector viruses were able to infect dog and rat cells. Bovine cells were infected only after their cocultivation with the retroviral packaging cell lines producing murine leukaemia virus vectors, perhaps as a result of a low

The foamy virus Pol protein is translated independently from Gag using a separate mRNA. Thus, in contrast to orthoretroviruses no Gag-Pol precursor protein is synthesized. Only the integrase domain is cleaved off from Pol resulting in a mature reverse transcriptase harboring the protease domain at the N-terminus (PR-RT). Although the homology between the PR-RTs from simian foamy virus from macaques (SFVmac) and the prototype foamy virus (PFV), probably originating from chimpanzee, exceeds 90%, several differences in the biophysical and biochemical properties of the two enzymes have been reported (i.e. SFVmac develops resistance to the nucleoside inhibitor azidothymidine (AZT) whereas PFV remains AZT sensitive even if the resistance mutations from SFVmac PR-RT are introduced into the PFV PR-RT gene). Moreover, contradictory data on the monomer/dimer status of the foamy virus protease have been published. We set out to purify and directly compare the monomer/dimer status and the enzymatic behavior of the

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The newly formed stable of NXT known as the Undisputed Era reveals what message they have for the rest of NXT. Click to read the full news