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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
24
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pubmed:dateCreated |
2010-11-24
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pubmed:abstractText |
Hepatitis C virus (HCV) nonstructural protein 5A (NS5A) exhibits a preference for G/U-rich RNA in vitro. Biological analysis of the NS5A RNA-binding activity and its target sites in the genome will be facilitated by a description of the NS5A-RNA complex. We demonstrate that the C-4 carbonyl of the uracil base and, by inference, the C-6 carbonyl of the guanine base interact with NS5A. U-rich RNA of 5 to 6 nucleotides (nt) is sufficient for high-affinity binding to NS5A. The minimal RNA-binding domain of NS5A consists of residues 2005 to 2221 (referred to as domain I-plus). This region of the protein includes the amino-terminal domain I as well as the subsequent linker that separates domains I and II. This linker region is the site of adaptive mutations. U-rich RNA-binding activity is not observed for an NS5A derivative containing only residues 2194 to 2419 (domains II and III). Mass spectrometric analysis of an NS5A-poly(rU) complex identified domains I and II as sites for interaction with RNA. Dimerization of NS5A was demonstrated by glutaraldehyde cross-linking. This dimerization is likely mediated by domain I-plus, as dimers of this protein are trapped by cross-linking. Dimers of the domain II-III protein are not observed. The monomer-dimer equilibrium of NS5A shifts in favor of dimer when U-rich RNA is present but not when A-rich RNA is present, consistent with an NS5A dimer being the RNA-binding-competent form of the protein. These data provide a molecular perspective of the NS5A-RNA complex and suggest possible mechanisms for regulation of HCV and cellular gene expression.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-10390360,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-11110665,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-11744739,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-12773396,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-15294292,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-15339921,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-15709040,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-15902263,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-16126720,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-16166071,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-16356462,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-16943283,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-18045871,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-18243120,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-18548002,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-18718931,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-19019822,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-19244328,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-19553194,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-19628699,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-19812153,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-20410884,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-20450484,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-20547756,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-20592076,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-7769656,
http://linkedlifedata.com/resource/pubmed/commentcorrection/20926572-7999043
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Dec
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pubmed:issn |
1098-5514
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pubmed:author |
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pubmed:issnType |
Electronic
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pubmed:volume |
84
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
12480-91
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pubmed:dateRevised |
2011-7-28
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pubmed:meshHeading |
pubmed-meshheading:20926572-Binding Sites,
pubmed-meshheading:20926572-Biotinylation,
pubmed-meshheading:20926572-Blotting, Western,
pubmed-meshheading:20926572-Cross-Linking Reagents,
pubmed-meshheading:20926572-Glutaral,
pubmed-meshheading:20926572-Humans,
pubmed-meshheading:20926572-Peptide Fragments,
pubmed-meshheading:20926572-Protein Binding,
pubmed-meshheading:20926572-Protein Structure, Tertiary,
pubmed-meshheading:20926572-RNA, Viral,
pubmed-meshheading:20926572-RNA-Binding Proteins,
pubmed-meshheading:20926572-Spectrometry, Mass, Matrix-Assisted Laser...,
pubmed-meshheading:20926572-Viral Nonstructural Proteins
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pubmed:year |
2010
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pubmed:articleTitle |
Hepatitis C virus nonstructural protein 5A: biochemical characterization of a novel structural class of RNA-binding proteins.
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pubmed:affiliation |
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, 201 Althouse Laboratory, University Park, PA 16802, USA.
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pubmed:publicationType |
Journal Article,
Research Support, N.I.H., Extramural
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