Fingerprint Dive into the research topics of Variability of the ,sup,15,/sup,N chemical shift anisotropy in Escherichia coli ribonuclease H in solution. Together they form a unique fingerprint. ...
The HIV-1 genomic RNA reverse transcription is an essential step in the virus cycle carried out by the viral-coded reverse transcriptase (RT), which has two associated functions: the RNA- and DNA-dependent DNA polymerase (RDDP and DDDP) function and the ribonuclease H (RNase H) function. The RNase H function catalyzes the selective hydrolysis of the RNA strand of the RNA:DNA heteroduplex replication intermediate. The RT associated activities are both essential for HIV-1 replication and validated targets for drug development, but only the polymerase function has been widely investigated as drug target. In fact, either nucleoside or non-nucleoside RT inhibitors currently used in therapy act on the polymerase associated activity. In this review, we describe the compounds, reported up to today, which inhibit the HIV-1 RNase H function, their chemical structures, the structure-activity relationships and the mechanism of action ...
Journal Article: Structural Basis for the Inhibition of RNase H Activity of HIV-1 Reverse Transcriptase by RNase H Active Site-Directed Inhibitors ...
This article communicates our study to elucidate the molecular determinants of weak Mg(2+) interaction with the ribonuclease H (RNH) domain of HIV-1 reverse transcriptase in solution. As the interaction is weak (a ligand-dissociation constant >1 mM), nonspecific Mg(2+) interaction with the protein or interaction of the protein with other solutes that are present in the buffer solution can confound the observed Mg(2+)-titration data. To investigate these indirect effects, we monitored changes in the chemical shifts of backbone amides of RNH by recording NMR (1)H-(15)N heteronuclear single-quantum coherence spectra upon titration of Mg(2+) into an RNH solution. We performed the titration under three different conditions: (1) in the absence of NaCl, (2) in the presence of 50 mM NaCl, and (3) at a constant 160 mM Cl(-) concentration. Careful analysis of these three sets of titration data, along with molecular dynamics simulation data of RNH with Na(+) and Cl(-) ions, demonstrates two characteristic ...
(2014) Figiel, Nowotny. Nucleic Acids Research. RNases H participate in the replication and maintenance of genomic DNA. RNase H1 cleaves the RNA strand of RNA/DNA hybrids, and RNase H2 in addition hydrolyzes the RNA residue of RNA-DNA junctions. RNase H3 is structurally closely related to RNases ...
This is new I think. Structural and Inhibition Studies of the RNase H Function of XMRV Reverse Transcriptase Karen A. Kirby1,2, Bruno...
Myc-DDK-tagged ORF clone of Homo sapiens ribonuclease H2, subunit B (RNASEH2B), transcript variant 2 as transfection-ready DNA - 10 µg - OriGene - cdna clones
A Computational Model for Predicting RNase H Domain of Retrovirus. . Biblioteca virtual para leer y descargar libros, documentos, trabajos y tesis universitarias en PDF. Material universiario, documentación y tareas realizadas por universitarios en nuestra biblioteca. Para descargar gratis y para leer online.
Chemicals. PM02734 was obtained from PharmaMar, prepared as a 10 mg/mL stock solution in DMSO/ethanol (1:1), and kept at −20°C. The drug concentrations used for the experiments were in the range of those obtained after clinical administration of the compound. Syringomycin E was also supplied by PharmaMar and was dissolved in DMSO at 5 mg/mL. MMS was supplied by Fluka and SDS was obtained from Sigma.. Yeast strains and media. The 4,848 S. cerevisiae haploid deletion mutants in nonessential genes were obtained from Invitrogen. Yeast strains were grown at 30°C on yeast extract-peptone-dextrose (YPD) with geneticin (G418, Duchefa) or without geneticin when growing the wild-type strain.. Electron microscopy. Cells (2 × 108) were fixed in 3% glutaraldehyde for 1 h at room temperature and processed as described by Wonisch et al. ( 4). Finally, cells were resuspended in 500 μL of 100% Spurr resin and transferred to capsules where resin polymerized for 20 h at 60°C. Ultrathin sections were stained ...
SWISS-MODEL Template Library (SMTL) entry for 1hrh.1. CRYSTAL STRUCTURE OF THE RIBONUCLEASE H DOMAIN OF HIV-1 REVERSE TRANSCRIPTASE
3O3G: Crystal Structures of RNase H2 in Complex with Nucleic Acid Reveal the Mechanism of RNA-DNA Junction Recognition and Cleavage.
TY - JOUR. T1 - Drugging the R-loop interactome. T2 - RNA-DNA hybrid binding proteins as targets for cancer therapy. AU - Boros-Oláh, Beáta. AU - Dobos, Nikoletta. AU - Hornyák, Lilla. AU - Szabó, Zoltán. AU - Karányi, Zsolt. AU - Halmos, Gábor. AU - Roszik, Jason. AU - Székvölgyi, Lóránt. PY - 2019/12. Y1 - 2019/12. N2 - Unravelling the origin of genetic alterations from point mutations to chromosomal rearrangements was greatly enhanced by the discovery of RNA-DNA hybrids (R-loops) that behave as hotspots of genomic instability in a variety of organisms. Current models suggest that uncontrolled R-loops are a hazard to genome integrity, therefore, identifying proteins that are involved in recognising and signalling R-loop structures are of key importance. Herein we analysed key RNA-DNA hybrid binding proteins in humans taking advantage of large-scale gene expression, survival rate, and drug-sensitivity data from cancer genomics databases. We show that expression of RNA-DNA hybrid ...
HIV-1 invert transcriptase (RT) catalyzes the conversion of genomic RNA into cDNA. comprehensive genetic evaluation of RT dimerization and really should make feasible the speedy screening process of potential inhibitors of the essential procedure. The HIV type 1 (HIV-1) invert transcriptase (RT) is necessary for the transformation of genomic RNA into double-stranded proviral DNA, catalyzed with the RNA- and DNA-dependent polymerase and ribonuclease H actions from the enzyme. HIV-1 RT can be an asymmetric dimer produced with the association of p51 and p66 polypeptides, that are cleaved from a big Pr160GagPol precursor with the viral protease during virion set up. p51 contains similar N-terminal sequences MK 3207 HCl manufacture as p66, but does not have the C-terminal ribonuclease H (RNase H) site (1). The framework of HIV-1 RT continues to be elucidated by x-ray crystallography in a MK 3207 HCl manufacture number of configurations, which includes MK 3207 HCl manufacture unliganded MK 3207 HCl ...
Ribonuclease H1, Ribonuclease H1, 5-R(*GP*GP*AP*GP*UP*GP*CP*GP*AP*CP*AP*CP*CP*UP*GP*AP*UP*UP*CP*C)-3), 5-D(*DGP*DGP*DAP*DAP*DTP*DCP*DAP*DGP*DGP*DTP*DGP*DTP*DCP*DGP*DCP*DAP*DCP*DTP*DCP*DT)-3 ...
The GScript RTase is recombinant M-MLV RTase expressed in E. coli and purified to homogeneity. It has lower RNase H activity and high thermal stability. The enzyme is widely used to synthesize first-strand cDNA at temperatures up to 55°C with increased specificity, higher yields of cDNA, and more full-length product than other reverse transcriptases. It can generate cDNA from 100 bp to 12 Kb ...
Callaghan, A. J.; Aurikko, J. P.; Illag, L. L.; Grossmann, G.; Chandran, V.; Kühnel, K.; Poljak, L.; Carpousis, A. J.; Robinson, C. V.; Symmons, M. F. et al.; Luisi, B. F.: Studies of the RNA degradosome-organizing domain of the Escherichia coli ribonuclease RNase E. Journal of Molecular Biology 340 (5), S. 965 - 979 (2004 ...
Mouse polyclonal Ribonuclease H2, subunit A antibody validated for WB, ICC/IF and tested in Human. Referenced in 2 publications. Immunogen corresponding to…
P32 appears to interact with the N-terminal duplex binding domain of RNase H1.(A) Expression and purification of RNase H1 deletion mutants. Left panel: Schemati
Ribonuclease H2 subunit A, also known as RNase H2 subunit A, is an enzyme that in humans is encoded by the RNASEH2A gene. The protein encoded by this gene is a component of the heterotrimeric type II ribonuclease H enzyme (RNaseH2). The other two subunits are the non-catalytic RNASEH2B and RNASEH2C. RNaseH2 is the major source of ribonuclease H activity in mammalian cells and endonucleolytically cleaves ribonucleotides. It is predicted to remove Okazaki fragment RNA primers during lagging strand DNA synthesis and to excise single ribonucleotides from DNA-DNA duplexes. Mutations in this gene cause Aicardi-Goutieres syndrome (AGS), an autosomal recessive neurological disorder characterized by progressive microcephaly and psychomotor retardation, intracranial calcifications, elevated levels of interferon-alpha and white blood cells in the cerebrospinal fluid. GRCh38: Ensembl release 89: ENSG00000104889 - Ensembl, May 2017 GRCm38: Ensembl release 89: ENSMUSG00000052926 - Ensembl, May 2017 "Human ...
TY - JOUR. T1 - Characterization of RNase HII substrate recognition using RNase HII-argonaute chimaeric enzymes from Pyrococcus furiosus. AU - Kitamura, Sayaka. AU - Fujishima, Kosuke. AU - Sato, Asako. AU - Tsuchiya, Daisuke. AU - Tomita, Masaru. AU - Kanai, Akio. PY - 2010/3/15. Y1 - 2010/3/15. N2 - RNase H (ribonuclease H) is an endonuclease that cleaves the RNA strand of RNA-DNA duplexes. It has been reported that the three-dimensional structure of RNase H is similar to that of the PIWI domain of the Pyrococcus furiosus Ago (argonaute) protein, although the two enzymes share almost no similarity in their amino acid sequences. Eukaryotic Ago proteins are key components of the RNA-induced silencing complex and are involved in microRNA or siRNA (small interfering RNA) recognition. In contrast, prokaryotic Ago proteins show greater affinity for RNA-DNA hybrids than for RNA-RNA hybrids. Interestingly, we found that wild-type Pf-RNase HII (P. furiosus, RNase HII) digests RNA-RNA duplexes in the ...
cansSAR 3D Structure of 6DPN_A | CRYSTAL STRUCTURE OF BACILLUS HALODURANS RIBONUCLEASE H1 E188A IN COMPLEX WITH AN RNA/DNA HYBRID: REACTION IN 2 MM MG2+ AND 200 MM K+ FOR 200 S AT 21 C | 6DPN
Electrospray ionization mass spectrometry (ESI-MS) conditions were optimized for simultaneous observation of a bimolecular qDNA and a Watson-Crick base-paired duplex DNA/RNA hybrid. The DNA sequence used was telomeric DNA, and the RNA contained the template for telomerase-mediated telomeric DNA synthesis. Addition of RNA to the quadruplex DNA (qDNA) resulted in formation of the duplex DNA/RNA hybrid. Melting profiles obtained using circular dichroism spectroscopy confirmed that the DNA/RNA hybrid exhibited greater thermal stability than the bimolecular qDNA in solution. Binding of a 13-substituted berberine (1) derivative to the bimolecular qDNA stabilized its structure as evidenced by an increase in its stability in the mass spectrometer, and an increase in its circular dichroism (CD) melting temperature of 10°C. The DNA/RNA hybrid did not bind the ligand extensively and its thermal stability was unchanged in the presence of (1). The qDNA-ligand complex resisted unfolding in the presence of excess RNA
Hits in gapped BLAST to ribonuclease HII sequences; e.g. residues 103-311 are 33% similar to RNH2_STRPN; residues 92-313 are 27% similar to (AE001275) Ribonuclease HII of Chlamydia trachomatis; and residues 103-304 are 23% similar to (AB012613) ribonuclease HII of Pyrococcus kodakaraensis ...
Title: Alabel-free and enzyme-free signal amplification strategy for a sensitive RNase Hactivity assay Author: Chang Yeol Lee,‡Hyowon Jang,‡ Ki Soo Park* and Hyun Gyu Park* (‡: equal contribution) Journal: Nanoscale 2017, 9, 16149-16153 Abstra
Title: Alabel-free and enzyme-free signal amplification strategy for a sensitive RNase Hactivity assay Author: Chang Yeol Lee,‡Hyowon Jang,‡ Ki Soo Park* and Hyun Gyu Park* (‡: equal contribution) Journal: Nanoscale 2017, 9, 16149-16153 Abstra
RNase H2B is a non-catalytic subunit of RNase H2, which removes both misincorporated ribonucleotides and R-loops from DNA. RNase H2B contains a PIP-box PCNA interaction motif and is responsible for nuclear localisation and interaction of RNase H2 with the DNA replication and repair machinery. We show that RNase H2 subunits are differentially expressed in several cancers. In colorectal cancer (CRC) tissues and cell lines, about 20% of cases have high RNase H2B expression, which correlates with decreased disease-free survival. We have generated RNase H2B- overexpressing CRC cells lines and show that RNase H2B overexpression reduces replication fork stalling and increases cell survival in response to replication stress ...
Rnaseh1: | | | Ribonuclease H1 | | | | |||| ... World Heritage Encyclopedia, the aggregation of the largest online encyclopedias available, and the most definitive collection ever assembled.
RNA interference (RNAi) is a post-transcriptional gene-silencing process that occurs in many eukaryotic organisms upon intracellular exposure to double-stranded RNA. Argonaute 2 (Ago2) protein is the catalytic engine of mammalian RNAi. It contains a PIWI domain that is structurally related to RNases H and possibly shares with them a two-metal-ion catalysis mechanism. Here we describe the expression in E. coli of mouse Ago2 and testing of its enzymatic activity in a RISC assay, i.e., for the ability to cleave a target RNA in a single position specified by a complementary small interfering RNA (siRNA). The results show that the enzyme can load the siRNA and cleave the complementary RNA in absence of other cellular factors, as described for human Ago2. It was also found that mutation of Arg669, a residue previously proposed to be involved in substrate and/or B metal ion binding, doesnt affect the enzymatic activity, suggesting that this residue doesnt belong to the active site.
Протеин кодиран овим геном је компонента хетеротримерне рибонуклеазе Х типа II (RNAseH2). RNAseH2 је главни извор рибонуклеазне Х активности у ћелијама сисара. Она ендонуклеолитички пресеца рибонуклеотид е Сматра се да уклања Оказакијев фрагмент РНК прајмера током синтезе заостајућег ланца ДНК и да исеца појединачне рибонуклеотиди из ДНК-ДНК дуплекса[1]. ...
R-loops at tRNA genes affect pre-tRNA synthesis in strains lacking topoisomerase and RNase H activities.A-E: Wild-type strain BY4741 (WT) and isogenic mutant
Robertson, H.D., Altman, S. and Smith, J.D. (1972). "Purification and properties of a specific Escherichia coli ribonuclease which cleaves a tyrosine transfer ribonucleic acid presursor". J. Biol. Chem. 247: 5243-5251. PMID 4560501. ...
Catalyzes the formation of a phosphodiester bond between juxtaposed 5´ phosphate and 3´ hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end and cohesive end termini as well as repair single stranded nicks in duplex DNA and some DNA/RNA hybrids. T4 DNA ligase will seal nicks for these DNA substrates.
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摘 要:上皮细胞转分化现象及其与疾病发生发展的关系,近年已成为细胞生物学、免疫学等多学科关注的聚焦点。转分化作为细胞分化发育的基本生物学现象,存在于机体诸多生理病理过程,也受表观遗传学的调控。相对于经典遗传学而言,表观遗传学作为一门新兴学科,其为生物体的基因表达调控及遗传现象提供了新的理论阐释。现知,DNA 甲基化、组蛋白修饰及非编码RNA 等均可导致上皮细胞基因发生表观遗传改变,与上皮细胞转分化的发生发展密切相关,并在该过程中发挥重要的调控作用。进一步阐明细胞转分化的分子基础及其表观遗传学调控机制,将有助于认识生命现象基本过程,并可为炎症性疾病、自身免疫病、器官纤维化,以及肿瘤发生与转移等机制的研究与防治,提供新的思路和应对策略。对上皮细胞转分化与表观遗传学调控关系作一简述 ...
Highly purified RNase H (RNA·DNA hybrid ribonucleotidohydrolase, EC 3.1.4.34) from calf thymus was used to specifically remove the poly(A) sequences of purified rabbit globin mRNA after its hybridization with poly(dT). The deadenylylated globin mRNA was repurified by a one-step procedure including a nitrocellulose column. The poly(A) size and the content of unmodified mRNA were determined by hybridization with [(3)H]-poly(U), and it could be shown that the RNase H digestion method effectively removes this terminal poly(A) sequence. No difference in activity was found between mRNAs with and without poly(A) to initiate, elongate, terminate, and release newly synthesized globin chains in exogenous-mRNA-dependent, cell-free, protein-synthesizing systems from wheat embryo, ascites Krebs II cells, and rat liver. Furthermore, poly(A)-free globin mRNA competed with the same efficiency as authentic globin mRNA against chick ovalbumin mRNA when translated under total mRNA saturation conditions. It is ...
The chronic engagement of the Ly49H activating receptor with m157 results in the impairment of NK cell function in vitro and in vivo (18, 19). In the studies described in this article, we demonstrate that NK cell impairment resulting from sustained engagement of Ly49H activating receptor in vivo requires signaling through the DAP12 adaptor molecule and is independent of DAP10. Additionally, we demonstrate that Ly49H-mediated NK cell tolerance can be established in mature NK cells and that the Ly49H-mediated NK cell tolerance is reversible in vivo.. The use of an in vivo transgenic mouse model offers advantages over in vitro models to study activation receptor-induced NK cell hyporesponsiveness. Primarily, NK cell activation assays in these experiments were performed on freshly isolated NK cells without the need for additional cytokines necessary for growing NK cells in vitro. Although it is necessary to use cytokines to grow in vitro, it is unclear whether these lymphokine-activated killer cells ...
This gene encodes an endonuclease that specifically degrades the RNA of RNA-DNA hybrids and is necessary for DNA replication and repair. This enzyme is present in both mitochondria and nuclei, which are resulted from translation of a single mRNA with two in-frame initiation start codons. The use of the first start codon produces the mitochondrial isoform and the use of the second start codon produces the nuclear isoform. The production of the mitochondrial isoform is modulated by an upstream open reading frame (uORF) which overlaps the first initiation start codon in human. An alternately spliced transcript variant has been found which encodes a shorter isoform. This gene has three pseudogenes; two of them are at different locations of chromosome 17 and one of them is on chromosome 1q32.2. [provided by RefSeq, Sep 2014 ...
Early post-infection, the reverse transcriptase converts the viral RNA genome into double-stranded viral DNA. The RNase H domain of the reverse transcriptase performs two functions. It degrades the RNA template and specifically removes the RNA primer from the RNA/DNA hybrid. Following nuclear import, the integrase catalyzes the insertion of the linear, double-stranded viral DNA into the host cell chromosome. Endogenous Pol proteins may have kept, lost or modified their original function during evolution (By similarity ...
TY - JOUR. T1 - NMR structure of an (-L-LNA:RNA hybrid: structural implications for RNase H recognition. AU - Nielsen, Jakob Toudahl. AU - Stein, Paul C.. AU - Petersen, M.. PY - 2003. Y1 - 2003. U2 - 10.1093/nar/gkg800. DO - 10.1093/nar/gkg800. M3 - Journal article. VL - 31. SP - 5858. EP - 5867. JO - Nucleic Acids Research. JF - Nucleic Acids Research. SN - 0305-1048. ER - ...
We follow expression of energy bypass genes, relative to other genes, under variable growth conditions and stressful challenges. One example is a collaborative effort to understand the molecular impact of switches in nitrogen nutrition, and associated pH changes.. By modifying genes for the mitochondrial NAD(P)H dehydrogenases we can influence the whole cell NAD(P)H levels, and directly assess how the cellular NAD(P)H pools control other processes. We have correlated transgenically induced changes in cellular NAD(P)(H) levels to changes in development. We now investigate redox responses to the environment, in particular NAD(P)H-mediated defences enabling plants to cope with biotic and abiotic stress. A particular interest is to analyse if energy bypasses counteract metabolic fluctuations induced by external stress.. ...
We have examined the antisense potency of the hybrid duplexes of fully-matched 3-, 5 and interior-chromophore tethered antisense oligos (AON) and three target RNAs (11mer and two 17mers) against RNase H, and found them to be better substrates compared t. ...
Human RNASEH2B full-length ORF ( AAH36744.1, 1 a.a. - 331 a.a.) recombinant protein with GST-tag at N-terminal. (H00079621-P01) - Products - Abnova
Human RNASEH2A full-length ORF ( AAH11748, 1 a.a. - 299 a.a.) recombinant protein with GST-tag at N-terminal. (H00010535-P01) - Products - Abnova
Page contains details about RNA-DNA hybrid origami barcode-like nanostructure . It has composition images, properties, Characterization methods, synthesis, applications and reference articles : nano.nature.com
strong-stop DNA: first species made during viral DNA synthesis containing sequences representing both the 5- & 3- end of the viral genome; initiated near the 3-end of the genome & copies 3 & 5 genomic sequences from (-) DNA
Lucigen strives to provide life scientists with the highest quality products and services for RNA/DNA amplification, cloning, next gen sequencing, and protein expression. Experience outstanding performance with time-saving convenience at an exceptional price.
i REAL-TIME PCR: FROM THEORY TO PRACTICE ii Put the odds in your favor with SuperScript RT. Engineered to be RNase H and incredibly thermostable, SuperScript III RT delivers robust first-strand synthesis
Converting the single-stranded retroviral RNA into integration-competent double-stranded DNA is achieved through a multi-step process mediated by the virus-coded reverse transcriptase (RT). With the exception that it is restricted to an intracellular life cycle, replication of the Saccharomyces cerevisiae long terminal repeat (LTR)-retrotransposon Ty3 genome is guided by equivalent events that, while generally similar, show many unique and subtle differences relative to the retroviral counterparts. Until only recently, our knowledge of RT structure and function was guided by a vast body of literature on the human immunodeficiency virus (HIV) enzyme. Although the recently-solved structure of Ty3 RT in the presence of an RNA/DNA hybrid adds little in terms of novelty to the mechanistic basis underlying DNA polymerase and ribonuclease H activity, it highlights quite remarkable topological differences between retroviral and LTR-retrotransposon RTs. The theme of overall similarity but distinct ...
Hepatitis B computer virus (HBV) synthesizes its DNA genome through reverse transcription which is catalyzed by viral polymerase (Pol). viral reverse transcription. Southern blot analysis showed that three mutants (R703A D777A and R781A mutants) yielded significantly reduced amounts of viral DNAs. However none of these mutants were defective in RNA encapsidation. The data indicated that in the R703A and D777A mutants minus-strand DNA synthesis was incomplete due to loss of catalytic activity of RNase H. In contrast in the R781A mutant the minus-strand DNA synthesis was near complete to some extent while the plus-strand DNA synthesis (i.e. relaxed circular DNA) was severely impaired due to the defect in RNase H activity. Overall our analysis revealed that three charged residues of the HBV Pol RNase H domain name contribute to the catalysis of RNase H in removing the RNA template but not in the RNA encapsidation. INTRODUCTION Hepatitis B computer virus (HBV) the prototypic member of the ...
Gene Tools makes Morpholino antisense oligos. Morpholino oligos bind to complementary RNA and get in the way of processes; they can knock down gene expression, modify RNA splicing or inhibit miRNA activity and maturation. Morpholinos are the premier knockdown tools used in developmental biology labs, the best RNA-blocking reagents for cells in culture and, as Vivo-Morpholinos, the most specific delivery-enhanced oligos available for other animal models. We are the sole commercial manufacturer selling research quantities of Morpholinos world-wide. Morpholino oligos are short chains of about 25 Morpholino subunits. Each subunit is comprised of a nucleic acid base, a morpholine ring and a non-ionic phosphorodiamidate intersubunit linkage. Morpholinos do not degrade their RNA targets, but instead act via an RNAse H-independent steric blocking mechanism. They are completely stable in cells, uncut by nucleases. With their requirement for greater complementarity with their target RNAs, Morpholinos are ...