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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
4
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pubmed:dateCreated |
1995-4-10
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pubmed:abstractText |
The selective encapsidation of retroviral RNA requires sequences in the Gag protein, as well as a cis-acting RNA packaging signal (psi site) near the 5' end of the genomic transcript. Gag protein of human immunodeficiency virus type 1 (HIV-1) has recently been found to bind specifically to the HIV-1 psi element in vitro. Here we report studies aimed at mapping features within the genetically defined psi locus that are required for binding of HIV-1 Gag or of its processed nucleocapsid derivative. The full-length HIV-1 Gag (p55) and nucleocapsid (p15) sequences were expressed as glutathione S-transferase (GST) fusion proteins in Escherichia coli. In a gel shift assay containing excess competitor tRNA, affinity-purified GST-p15 and GST-p55 proteins bound to a 206-nucleotide psi RNA element spanning the major splice donor and gag start codons but did not bind to antisense psi transcripts. Quantitative filter-binding assays revealed that both GST-p55 and GST-p15 bound to this RNA sequence with identical affinities (apparent Kd congruent to 5 x 10(-8) M), indicating that all major determinants of psi binding affinity reside within the nucleocapsid portion of Gag. Chemical and RNase accessibility mapping, coupled with computerized sequence analysis, suggested a model for psi RNA structure comprising four independent stem-loops. Filter-binding studies revealed that RNAs corresponding to three of these hypothetical stem-loops can each function as a independent Gag binding site and that each is bound with approximately fourfold-lower apparent affinity than the full-length psi locus. Interaction of Gag with these regions is likely to play a major role in directing HIV-1 RNA encapsidation in vivo.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-1309906,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-1378506,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-1560523,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-1585635,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-2109098,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-2124274,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-2191147,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-3012114,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-3033290,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-3039161,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-3418786,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-3502707,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-3989912,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-434465,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-6253670,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-6310558,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-6310871,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-6313966,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-6678608,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8035501,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8156990,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8189516,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8197162,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8207823,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8218211,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8230441,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8248806,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8313915,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8371356,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8380070,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8429553,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8443588,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8475087,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8506369,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7884856-8510213
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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 |
Apr
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pubmed:issn |
0022-538X
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
69
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pubmed:geneSymbol |
gag
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
2101-9
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
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pubmed:year |
1995
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pubmed:articleTitle |
RNA secondary structure and binding sites for gag gene products in the 5' packaging signal of human immunodeficiency virus type 1.
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pubmed:affiliation |
Department of Pathology, University of California, San Francisco 94143-0506.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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