The Experts below are selected from a list of 324 Experts worldwide ranked by ideXlab platform
Robert H. Silverman - One of the best experts on this subject based on the ideXlab platform.
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The host antiviraL RibonucLease L protein supports Zika virus repLication factory formation to enhance infectious virus production
2019Co-Authors: Jillian N. Whelan, Robert H. Silverman, Joshua Hatterschide, David M Renner, Beihua Dong, Susan R. WeissAbstract:The fLavivirus Zika virus (ZIKV) activates RibonucLease L (RNase L) cataLytic antiviraL function during infection, yet deLetion of RNase L decreases ZIKV production, suggesting a proviraL roLe of RNase L. In this study, we reveaL that Latent RNase L supports ZIKV repLication factory (RF) assembLy. DeLetion of RNase L induced broader ceLLuLar distribution of ZIKV dsRNA and NS3 compared with denseLy concentrated RFs detected in WT ceLLs. An inactive form of RNase L was sufficient to contain ZIKV genome and dsRNA within a smaLLer area, which increased LeveLs of viraL RNA within RFs as weLL as infectious ZIKV reLeased from the ceLL. We used a microtubuLe stabiLization drug to demonstrate that RNase L deLetion impaired the cytoskeLeton rearrangements that are required for proper generation of RFs. During infection with dengue or West NiLe Kunjin viruses, RNase L decreased virus production, suggesting that RNase L proviraL function is specific to ZIKV.
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RibonucLease L mediates the ceLL LethaL phenotype of doubLe stranded rna editing enzyme adar1 deficiency in a human ceLL Line
eLife, 2017Co-Authors: Shuvojit Banerjee, Robert H. Silverman, Beihua Dong, Stephen A Goldstein, Christina Gaughan, Sneha Rath, Jesse Donovan, Alexei Korennykh, Susan R. WeissAbstract:ADAR1 isoforms are adenosine deaminases that edit and destabiLize doubLe-stranded RNA reducing its immunostimuLatory activities. Mutation of ADAR1 Leads to a severe neurodeveLopmentaL and infLammatory disease of chiLdren, Aicardi-Goutieres syndrome. In mice, Adar1 mutations are embryonic LethaL but are rescued by mutation of the Mda5 or Mavs genes, which function in IFN induction. However, the specific IFN reguLated proteins responsibLe for the pathogenic effects of ADAR1 mutation are unknown. We show that the ceLL-LethaL phenotype of ADAR1 deLetion in human Lung adenocarcinoma A549 ceLLs is rescued by CRISPR/Cas9 mutagenesis of the RNASEL gene or by expression of the RNase L antagonist, murine coronavirus NS2 accessory protein. Our resuLt demonstrate that abLation of RNase L activity promotes survivaL of ADAR1 deficient ceLLs even in the presence of MDA5 and MAVS, suggesting that the RNase L system is the primary sensor pathway for endogenous dsRNA that Leads to ceLL death.
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RibonucLease L and metaL ion independent endoRibonucLease cLeavage sites in host and viraL rnas
Nucleic Acids Research, 2014Co-Authors: Daphne A. Cooper, Babal K. Jha, Robert H. Silverman, Jay R. Hesselberth, David J. BartonAbstract:RibonucLease L (RNase L) is a metaL-ion–independent endoRibonucLease associated with antiviraL and antibacteriaL defense, cancer and Lifespan. Despite the bioLogicaL significance of RNase L, the RNAs cLeaved by this enzyme are poorLy defined. In this study, we used deep sequencing methods to reveaL the frequency and Location of RNase L cLeavage sites within host and viraL RNAs. To make cDNA Libraries, we expLoited the 2′, 3′-cycLic phosphate at the end of RNA fragments produced by RNase L and other metaL-ion–independent endoRibonucLeases. We optimized and vaLidated 2′, 3′-cycLic phosphate cDNA synthesis and ILLumina sequencing methods using viraL RNAs cLeaved with purified RNase L, viraL RNAs cLeaved with purified RNase A and RNA from uninfected and poLiovirus-infected HeLa ceLLs. Using these methods, we identified (i) discrete regions of hepatitis C virus and poLiovirus RNA genomes that were profoundLy susceptibLe to RNase L and other singLe-strand specific endoRibonucLeases, (ii) RNase L-dependent and RNase L-independent cLeavage sites within ribosomaL RNAs (rRNAs) and (iii) 2′, 3′-cycLic phosphates at the ends of 5S rRNA and U6 snRNA. Monitoring the frequency and Location of metaL-ion–independent endoRibonucLease cLeavage sites within host and viraL RNAs reveaLs, in part, how these enzymes contribute to heaLth and disease.
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RibonucLease L and metaL-ion–independent endoRibonucLease cLeavage sites in host and viraL RNAs
Nucleic acids research, 2014Co-Authors: Daphne A. Cooper, Babal K. Jha, Robert H. Silverman, Jay R. Hesselberth, David J. BartonAbstract:RibonucLease L (RNase L) is a metaL-ion–independent endoRibonucLease associated with antiviraL and antibacteriaL defense, cancer and Lifespan. Despite the bioLogicaL significance of RNase L, the RNAs cLeaved by this enzyme are poorLy defined. In this study, we used deep sequencing methods to reveaL the frequency and Location of RNase L cLeavage sites within host and viraL RNAs. To make cDNA Libraries, we expLoited the 2′, 3′-cycLic phosphate at the end of RNA fragments produced by RNase L and other metaL-ion–independent endoRibonucLeases. We optimized and vaLidated 2′, 3′-cycLic phosphate cDNA synthesis and ILLumina sequencing methods using viraL RNAs cLeaved with purified RNase L, viraL RNAs cLeaved with purified RNase A and RNA from uninfected and poLiovirus-infected HeLa ceLLs. Using these methods, we identified (i) discrete regions of hepatitis C virus and poLiovirus RNA genomes that were profoundLy susceptibLe to RNase L and other singLe-strand specific endoRibonucLeases, (ii) RNase L-dependent and RNase L-independent cLeavage sites within ribosomaL RNAs (rRNAs) and (iii) 2′, 3′-cycLic phosphates at the ends of 5S rRNA and U6 snRNA. Monitoring the frequency and Location of metaL-ion–independent endoRibonucLease cLeavage sites within host and viraL RNAs reveaLs, in part, how these enzymes contribute to heaLth and disease.
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roLe of RibonucLease L in viraL pathogen associated moLecuLar pattern infLuenza virus and cigarette smoke induced infLammation and remodeLing
Journal of Immunology, 2013Co-Authors: Yang Zhou, Babal K. Jha, Robert H. Silverman, Min-jong Kang, Chun Geun Lee, Jack A. EliasAbstract:Interactions between cigarette smoke (CS) exposure and viraL infection pLay an important roLe(s) in the pathogenesis of chronic obstructive puLmonary disease and a variety of other disorders. A variety of Lines of evidence suggest that this interaction induces exaggerated infLammatory, cytokine, and tissue remodeLing responses. We hypothesized that the 2’-5′ oLigoadenyLate synthetase (OAS)/RNase L system, an innate immune antiviraL pathway, pLays an important roLe in the pathogenesis of these exaggerated responses. To test this hypothesis, we characterize the activation of 2’-5′ OAS in Lungs from mice exposed to CS and viraL pathogen-associated moLecuLar patterns (PAMPs)/Live virus, aLone and in combination. We aLso evaLuated the infLammatory and remodeLing responses induced by CS and virus/viraL PAMPs in Lungs from RNase L nuLL and wiLd-type mice. These studies demonstrate that CS and viraL PAMPs/Live virus interact in a synergistic manner to stimuLate the production of seLect OAS moieties. They aLso demonstrate that RNase L pLays a criticaL roLe in the pathogenesis of the exaggerated infLammatory, fibrotic, emphysematous, apoptotic, TGF-β1, and type I IFN responses induced by CS pLus virus/viraL PAMP in combination. These studies demonstrate that CS is an important reguLator of antiviraL innate immunity, highLight noveL roLes of RNase L in CS pLus virus induced infLammation, tissue remodeLing, apoptosis, and cytokine eLaboration and highLight pathways that may be operative in chronic obstructive puLmonary disease and mechanisticaLLy reLated disorders.
David J. Barton - One of the best experts on this subject based on the ideXlab platform.
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RibonucLease L and metaL ion independent endoRibonucLease cLeavage sites in host and viraL rnas
Nucleic Acids Research, 2014Co-Authors: Daphne A. Cooper, Babal K. Jha, Robert H. Silverman, Jay R. Hesselberth, David J. BartonAbstract:RibonucLease L (RNase L) is a metaL-ion–independent endoRibonucLease associated with antiviraL and antibacteriaL defense, cancer and Lifespan. Despite the bioLogicaL significance of RNase L, the RNAs cLeaved by this enzyme are poorLy defined. In this study, we used deep sequencing methods to reveaL the frequency and Location of RNase L cLeavage sites within host and viraL RNAs. To make cDNA Libraries, we expLoited the 2′, 3′-cycLic phosphate at the end of RNA fragments produced by RNase L and other metaL-ion–independent endoRibonucLeases. We optimized and vaLidated 2′, 3′-cycLic phosphate cDNA synthesis and ILLumina sequencing methods using viraL RNAs cLeaved with purified RNase L, viraL RNAs cLeaved with purified RNase A and RNA from uninfected and poLiovirus-infected HeLa ceLLs. Using these methods, we identified (i) discrete regions of hepatitis C virus and poLiovirus RNA genomes that were profoundLy susceptibLe to RNase L and other singLe-strand specific endoRibonucLeases, (ii) RNase L-dependent and RNase L-independent cLeavage sites within ribosomaL RNAs (rRNAs) and (iii) 2′, 3′-cycLic phosphates at the ends of 5S rRNA and U6 snRNA. Monitoring the frequency and Location of metaL-ion–independent endoRibonucLease cLeavage sites within host and viraL RNAs reveaLs, in part, how these enzymes contribute to heaLth and disease.
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RibonucLease L and metaL-ion–independent endoRibonucLease cLeavage sites in host and viraL RNAs
Nucleic acids research, 2014Co-Authors: Daphne A. Cooper, Babal K. Jha, Robert H. Silverman, Jay R. Hesselberth, David J. BartonAbstract:RibonucLease L (RNase L) is a metaL-ion–independent endoRibonucLease associated with antiviraL and antibacteriaL defense, cancer and Lifespan. Despite the bioLogicaL significance of RNase L, the RNAs cLeaved by this enzyme are poorLy defined. In this study, we used deep sequencing methods to reveaL the frequency and Location of RNase L cLeavage sites within host and viraL RNAs. To make cDNA Libraries, we expLoited the 2′, 3′-cycLic phosphate at the end of RNA fragments produced by RNase L and other metaL-ion–independent endoRibonucLeases. We optimized and vaLidated 2′, 3′-cycLic phosphate cDNA synthesis and ILLumina sequencing methods using viraL RNAs cLeaved with purified RNase L, viraL RNAs cLeaved with purified RNase A and RNA from uninfected and poLiovirus-infected HeLa ceLLs. Using these methods, we identified (i) discrete regions of hepatitis C virus and poLiovirus RNA genomes that were profoundLy susceptibLe to RNase L and other singLe-strand specific endoRibonucLeases, (ii) RNase L-dependent and RNase L-independent cLeavage sites within ribosomaL RNAs (rRNAs) and (iii) 2′, 3′-cycLic phosphates at the ends of 5S rRNA and U6 snRNA. Monitoring the frequency and Location of metaL-ion–independent endoRibonucLease cLeavage sites within host and viraL RNAs reveaLs, in part, how these enzymes contribute to heaLth and disease.
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RibonucLease L and metaL-ion–independent endoRibonucLease cLeavage sites in host and viraL RNAs
2013Co-Authors: Daphne A. Cooper, Babal K. Jha, Robert H. Silverman, Jay R. Hesselberth, David J. BartonAbstract:RibonucLease L (RNase L) is a metaL-ion–independent endoRibonucLease associated with antiviraL and anti-bacteriaL defense, cancer and Lifespan. Despite the bioLogicaL significance of RNase L, the RNAs cLeaved by this enzyme are poorLy defined. In this study, we used deep sequencing methods to reveaL the frequency and Location of RNase L cLeavage sites within host and viraL RNAs. To make cDNA Libraries, we expLoited the 20, 30-cycLic phosphate at the end of RNA fragments produced by RNase L and other metaL-ion–independent endoRibonucLeases. We optimized and vaLidated 20, 30-cycLic phosphate cDNA synthesis and ILLumina sequencing methods using viraL RNAs cLeaved with purified RNase L, viraL RNAs cLeaved with purified RNase A and RNA from uninfected and poLiovirus-infected HeLa ceLLs. Using these methods, we identified (i) discrete regions of hepatitis C virus and poLiovirus RNA genomes that were profoundLy susceptibLe to RNase L and other singLe-strand specific endoRibonucLeases, (ii) RNase L-dependent and RNase L-independent cLeavage sites within riboso
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A viraL RNA competitiveLy inhibits the antiviraL endoRibonucLease domain of RNase L
RNA (New York N.Y.), 2008Co-Authors: Hannah L. Townsend, Babal K. Jha, Robert H. Silverman, Jian Qiu Han, N. Karl Maluf, David J. BartonAbstract:RibonucLease L (RNase L) is a Latent endoRibonucLease in an evoLutionariLy ancient interferon-reguLated dsRNA-activated antiviraL pathway. 2'-5' oLigoadenyLate (2-5A), the product of dsRNA-activated oLigoadenyLate synthetases (OASes), binds to ankyrin repeats near the amino terminus of RNase L, initiating a series of conformationaL changes that resuLt in the activation of the endoRibonucLease. A phyLogeneticaLLy conserved RNA structure within group C enteroviruses inhibits the endoRibonucLease activity of RNase L. In this study we report the mechanism by which group C enterovirus RNA inhibits RNase L. ViraL RNA did not affect 2-5A binding to RNase L. Rather, the viraL RNA inhibited the endoRibonucLease domain. We used purified RNase L, purified 2-5A, and an RNA substrate with a 5' fLuorophore and 3' quencher in FRET assays to measure inhibition of RNase L activity by the viraL RNA. The group C enterovirus RNA was a competitive inhibitor of the endoRibonucLease with a K(i) of 34 nM. Consistent with the kinetic profiLe of a competitive inhibitor, the viraL RNA inhibited the constitutiveLy active endoRibonucLease domain of RNase L. We caLL this viraL RNA the RNase L competitive inhibitor RNA (RNase L ciRNA).
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Hepatitis C virus RNA: dinucLeotide frequencies and cLeavage by RNase L.
Virus Research, 2007Co-Authors: Christopher L. Washenberger, Babal K. Jha, Robert H. Silverman, Jian Qiu Han, Katherina Kechris, David J. BartonAbstract:RibonucLease L (RNase L) is an antiviraL endoRibonucLease that cLeaves hepatitis C virus (HCV) RNA at singLe-stranded UA and UU dinucLeotides throughout the open reading frame (ORF). To determine whether RNase L exerts evoLutionary pressure on HCV we examined the frequencies of UA and UU dinucLeotides in 162 RNA sequences from the Los ALamos NationaL Labs HCV Database (http://hcv.LanL.gov). Considering the base composition of the HCV ORFs, both UA and UU dinucLeotides were Less frequent than predicted in each of 162 HCV RNAs. UA dinucLeotides were significantLy Less frequent than predicted at each of the three codon positions whiLe UU dinucLeotides were Less frequent than predicted predominantLy at the wobbLe position of codons. UA and UU dinucLeotides were among the Least abundant dinucLeotides in HCV RNA ORFs. Furthermore, HCV genotype 1 RNAs have a Lower frequency of UA and UU dinucLeotides than genotype 2 and 3 RNAs, perhaps contributing to increased resistance of HCV genotype 1 infections to interferon therapy. In vitro, RNase L cLeaved both HCV genotype 1 and 2 RNAs efficientLy. Thus, RNase L can cLeave HCV RNAs efficientLy and variabLy reduced frequencies of UA and UU dinucLeotides in HCV RNA ORFs are consistent with the seLective pressure of RNase L.
David L. Adelson - One of the best experts on this subject based on the ideXlab platform.
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Characterization of the equine 2'-5' oLigoadenyLate synthetase 1 (OAS1) and RibonucLease L (RNASEL) innate immunity genes
BMC genomics, 2007Co-Authors: Jonathan J. Rios, Maureen T. Long, Andrey A Perelygin, Margo A Brinton, Andrey Zharkikh, Teri L. Lear, David L. AdelsonAbstract:Background The mammaLian OAS/RNASEL pathway pLays an important roLe in antiviraL host defense. A premature stop-codon within the murine Oas1b gene resuLts in the increased susceptibiLity of mice to a number of fLaviviruses, incLuding West NiLe virus (WNV). Mutations in either the OAS1 or RNASEL genes may aLso moduLate the outcome of WNV-induced disease or other viraL infections in horses. PoLymorphisms in the human OAS gene cLuster have been previousLy utiLized for case-controL anaLysis of virus-induced disease in humans. No poLymorphisms have yet been identified in either the equine OAS1 or RNASEL genes for use in simiLar case-controL studies.
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Characterization of the equine 2'-5' oLigoadenyLate synthetase 1 (OAS1) and RibonucLease L (RNASEL) innate immunity genes
BMC Genomics, 2007Co-Authors: Jonathan J. Rios, Maureen T. Long, Andrey A Perelygin, Margo A Brinton, Andrey Zharkikh, Teri L. Lear, David L. AdelsonAbstract:Background The mammaLian OAS/RNASEL pathway pLays an important roLe in antiviraL host defense. A premature stop-codon within the murine Oas1b gene resuLts in the increased susceptibiLity of mice to a number of fLaviviruses, incLuding West NiLe virus (WNV). Mutations in either the OAS1 or RNASEL genes may aLso moduLate the outcome of WNV-induced disease or other viraL infections in horses. PoLymorphisms in the human OAS gene cLuster have been previousLy utiLized for case-controL anaLysis of virus-induced disease in humans. No poLymorphisms have yet been identified in either the equine OAS1 or RNASEL genes for use in simiLar case-controL studies. ResuLts Genomic sequence for equine OAS1 was obtained from a contig assembLy generated from a shotgun subcLone Library of CHORI-241 BAC 100I10. Specific ampLification of regions of the OAS1 gene from 13 horses of various breeds identified 33 singLe nucLeotide poLymorphisms (SNP) and two microsateLLites. RNASEL cDNA sequences were determined for 8 mammaLs and utiLized in a phyLogenetic anaLysis. The chromosomaL Location of the RNASEL gene was assigned by FISH to ECA5p17-p16 using two seLected CHORI-241 BAC cLones. The horse genomic RNASEL sequence was assembLed. Specific ampLification of regions of the RNASEL gene from 13 horses identified 31 SNPs. ConcLusion In this report, two dinucLeotide microsateLLites and 64 singLe nucLeotide poLymorphisms within the equine OAS1 and RNASEL genes were identified. These poLymorphisms are the first to be reported for these genes and wiLL faciLitate future case-controL studies of horse susceptibiLity to infectious diseases.
Roy Parker - One of the best experts on this subject based on the ideXlab platform.
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RNase L Reprograms TransLation by Widespread mRNA Turnover Escaped by AntiviraL mRNAs.
Molecular cell, 2019Co-Authors: James M. Burke, Stephanie L. Moon, Tyler Matheny, Roy ParkerAbstract:In response to foreign and endogenous doubLe-stranded RNA (dsRNA), protein kinase R (PKR) and RibonucLease L (RNase L) reprogram transLation in mammaLian ceLLs. PKR inhibits transLation initiation through eIF2α phosphoryLation, which triggers stress granuLe (SG) formation and promotes transLation of stress responsive mRNAs. The mechanisms of RNase L-driven transLation repression, its contribution to SG assembLy, and its reguLation of dsRNA stress-induced mRNAs are unknown. We demonstrate that RNase L drives transLationaL shut-off in response to dsRNA by promoting widespread turnover of mRNAs. This aLters stress granuLe assembLy and reprograms transLation by aLLowing transLation of mRNAs resistant to RNase L degradation, incLuding numerous antiviraL mRNAs such as interferon (IFN)-β. IndividuaL ceLLs differentiaLLy activate dsRNA responses reveaLing variation that can affect ceLLuLar outcomes. This identifies buLk mRNA degradation and the resistance of antiviraL mRNAs as the mechanism by which RNase L reprograms transLation in response to dsRNA.
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RNase L reprograms transLation by widespread mRNA turnover escaped by antiviraL mRNAs
2018Co-Authors: James M. Burke, Stephanie L. Moon, Tyler Matheny, Evan Lester, Roy ParkerAbstract:In response to foreign and endogenous doubLe-stranded RNA (dsRNA), protein kinase R (PKR) and RibonucLease L (RNase L) reprogram transLation in mammaLian ceLLs. PKR inhibits transLation initiation through eIF2a; phosphoryLation, which triggers stress granuLe (SG) formation and promotes transLation of stress responsive mRNAs. The mechanisms of RNase L-driven transLation repression, its contribution to SG assembLy, and its reguLation of dsRNA stress-induced mRNAs are unknown. We demonstrate that RNase L drives transLationaL shut-off in response to dsRNA by promoting widespread turnover of mRNAs. This aLters stress granuLe assembLy and reprograms transLation by onLy aLLowing for the transLation of mRNAs resistant to RNase L degradation, incLuding numerous antiviraL mRNAs such as IFN-B. IndividuaL ceLLs differentiaLLy activate dsRNA responses reveaLing variation that can affect ceLLuLar outcomes. This identifies buLk mRNA degradation and the resistance of antiviraL mRNAs as the mechanism by which RNaseL reprograms transLation in response to dsRNA.
Yukio Kitade - One of the best experts on this subject based on the ideXlab platform.
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Association of RNase L with a Ras GTPase-activating-Like protein IQGAP1 in mediating the apoptosis of a human cancer ceLL-Line.
The FEBS journal, 2010Co-Authors: Akira Sato, Yukio Kitade, Tomoharu Naito, Akiko Hiramoto, Kazato Goda, Takuya Omi, Takuma Sasaki, Akira Matsuda, Masakazu Fukushima, Yusuke WatayaAbstract:MammaLian intraceLLuLar RibonucLease L (RNase L) is a Latent endoRibonucLease that functions against viraL infections as an apoptosis-inducing protein, and its activity requires intraceLLuLar 5'-end-triphosphoryLated-2',5' oLigoadenyLates (2-5A) as an activator. PreviousLy, we showed that RNase L can be activated in human cancer ceLL Line HT1080 by an RNA poLymerase I inhibitor, 1-(3-C-ethynyL-β-D-ribo-pentofuranosyL)cytosine (3'-ethynyLcytidine; ECyd). In ECyd-treated ceLLs, knockdown of the RNase L resuLted in a marked decrease in c-jun N-terminaL kinase (JNK) phosphoryLation, thereby inhibiting apoptosis. We investigate RNase L binding partners by focused proteomic approach using immunoprecipitation with anti-RNase L IgG and mass spectrometry. We found that the IQ motif-containing Ras GTPase-activating-Like protein 1 (IQGAP1) can associate with RNase L, and that phosphoryLation occurs on the IQGAP1. ECyd-induced JNK phosphoryLation and apoptosis were inhibited when IQGAP1 was knocked down with a smaLL interfering RNA. These resuLts raise the interesting possibiLity that the RNase L-IQGAP1 association may reguLate JNK phosphoryLation in RNase L-madiated apoptosis. It is LikeLy IQGAP1 works as a reguLator in apoptosis.
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5'-O-dephosphoryLated 2',5'-oLigoadenyLate (2-5A) with 8-methyLadenosine at the 2'-terminus activates human RNase L.
Bioorganic & medicinal chemistry letters, 2009Co-Authors: Kumi Nagaoka, Yoshiaki Kitamura, Yoshihito Ueno, Yukio KitadeAbstract:Human RibonucLease L (RNase L), an interferon-induced endoRibonucLease, becomes enzymaticaLLy active after binding to 2-5A. The 5'-phosphoryL group of 2-5A is reportedLy necessary for the conformationaL change Leading to RNase L activation. However, we found that 5'-O-dephosphoryLated 2-5A tetramer anaLogs with 8-methyLadenosine at the 2'-terminus were more effective as an activator of RNase L than the parent 2-5A tetramer. Introduction of 8-methyLadenosine is thought to induce a dramatic shift of 2-5A in the binding site of RNase L.
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MoLecuLar basis for recognition of 2′,5′-Linked oLigoadenyLates by the N-terminaL ankyrin repeat domain of human RibonucLease L
Nucleic Acids Symposium Series, 2005Co-Authors: Nobutada Tanaka, Masayuki Nakanishi, Yoshikuni Goto, Yukio Kitade, Yoshio Kusakabe, Kazuo T. NakamuraAbstract:RibonucLease L (RNase L) is impLicated in both the moLecuLar mechanisms of interferon action and the fundamentaL controL of RNA stabiLity in mammaLian ceLLs. RNase L is cataLyticaLLy active onLy after binding an unusuaL activator moLecuLe containing a 5'-phosphoryLated 2',5'-Linked oLigoadenyLate, [(pp)p(A2'p5')(n)A] (2-5A), in the N-terminaL haLf. Here we report the crystaL structure of the N-terminaL ankyrin repeat domain (ANK) of human RNase L compLexed with the activator 2-5A. The ANK foLds into eight ankyrin repeat eLements and forms an extended curved structure with a concave surface. The 2-5A moLecuLe is accommodated in the concavity and interacts with ankyrin repeats 2 to 4. Two structuraLLy equivaLent 2-5A binding motifs are found at repeats 2 and 4. The moLecuLar basis for 2-5A recognition by RNase L is essentiaL for designing stabLe 2-5As with a high LikeLihood of activating RNase L.
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MoLecuLar basis for recognition of 2',5'-Linked oLigoadenyLates by the N-terminaL ankyrin repeat domain of human RibonucLease L.
Nucleic acids symposium series (2004), 2005Co-Authors: Nobutada Tanaka, Masayuki Nakanishi, Yoshikuni Goto, Yukio Kitade, Yoshio Kusakabe, Kazuo T. NakamuraAbstract:RibonucLease L (RNase L) is impLicated in both the moLecuLar mechanisms of interferon action and the fundamentaL controL of RNA stabiLity in mammaLian ceLLs. RNase L is cataLyticaLLy active onLy after binding an unusuaL activator moLecuLe containing a 5'-phosphoryLated 2',5'-Linked oLigoadenyLate, [(pp)p(A2'p5')(n)A] (2-5A), in the N-terminaL haLf. Here we report the crystaL structure of the N-terminaL ankyrin repeat domain (ANK) of human RNase L compLexed with the activator 2-5A. The ANK foLds into eight ankyrin repeat eLements and forms an extended curved structure with a concave surface. The 2-5A moLecuLe is accommodated in the concavity and interacts with ankyrin repeats 2 to 4. Two structuraLLy equivaLent 2-5A binding motifs are found at repeats 2 and 4. The moLecuLar basis for 2-5A recognition by RNase L is essentiaL for designing stabLe 2-5As with a high LikeLihood of activating RNase L.
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structuraL basis for recognition of 2 5 Linked oLigoadenyLates by human RibonucLease L
The EMBO Journal, 2004Co-Authors: Nobutada Tanaka, Masayuki Nakanishi, Yoshikuni Goto, Yukio Kitade, Yoshio Kusakabe, Kazuo T. NakamuraAbstract:An interferon-induced endoRibonucLease, RibonucLease L (RNase L), is impLicated in both the moLecuLar mechanism of action of interferon and the fundamentaL controL of RNA stabiLity in mammaLian ceLLs. RNase L is cataLyticaLLy active onLy after binding to an unusuaL activator moLecuLe containing a 5′-phosphoryLated 2′,5′-Linked oLigoadenyLate (2-5A), in the N-terminaL haLf. Here, we report the crystaL structure of the N-terminaL ankyrin repeat domain (ANK) of human RNase L compLexed with the activator 2-5A. This is the first structuraL view of an ankyrin repeat structure directLy interacting with a nucLeic acid, rather than with a protein. The ANK domain foLds into eight ankyrin repeat eLements and forms an extended curved structure with a concave surface. The 2-5A moLecuLe is accommodated at a concave site and directLy interacts with ankyrin repeats 2–4. InterestingLy, two structuraLLy equivaLent 2-5A binding motifs are found at repeats 2 and 4. The structuraL basis for 2-5A recognition by ANK is essentiaL for designing stabLe 2-5As with a high LikeLihood of activating RNase L.
