Document Detail

Human immunodeficiency virus type 1 preferentially encapsidates genomic RNAs that encode Pr55(Gag): functional linkage between translation and RNA packaging.
MedLine Citation:
PMID:  11886257     Owner:  NLM     Status:  MEDLINE    
Full-length retroviral RNA serves as both messenger and genomic RNA. Therefore, an unspliced RNA could play both roles: viral mRNA could be bound in cis by the same Gag polyprotein that it produced, becoming a packaged genomic RNA. To test this possibility, we used in vivo packaging experiments which coexpressed wild-type NL4-3 RNA and NL4-3-based mutant RNA that, ideally, could not translate Gag. However, mutating the gag initiator produced a mutant (pNLX) that expressed a truncated Gag, Gag*, initiated at methionine 10 in the CA region (142 of Pr55(Gag)). Gag* can be rescued into virions by Gag and, as it contains the NC domain, could package RNA in cis. To eliminate NC and the CA dimerization domain, a nonsense mutation in CA at residue 99 was introduced into pNLX to produce pNLXX, which expresses an RNA that should only be packaged in trans. Cotransfection packaging experiments revealed that wild-type genomic RNA was packaged at an 8-fold greater level than NLXX RNA given equal expression of both RNAs. Experiments that varied the relative amounts of these RNAs in the cell found that the wild-type RNA was encapsidated with a packaging preference (i.e., the relative amount of this RNA in virions versus cells) of 6- to 13-fold over the NLXX RNA, showing that the NLXX RNA did not efficiently compete with NL4-3 RNA. These data suggest that the wild-type RNA's ability to express Pr55(Gag) and, by inference, actively translate Gag confers an advantage in packaging over the nearly identical NLXX RNA. In contrast, the NLX RNA competed with wild-type RNA at a 1-to-3 preference. This ratio is similar to the amounts of Gag* rescued by Gag, suggesting that the presence of Gag* assists in the encapsidation of NLX RNA. Together, our data link translation and particle formation to the packaging of viral RNA and support a model of cis packaging where nascent Gag proteins encapsidate their cognate RNA.
Dexter T K Poon; Elena N Chertova; David E Ott
Related Documents :
8673017 - A simple and efficient method for the isolation of total rna from the fission yeast sch...
1711537 - Expression of immunoreactive growth hormone in leukocytes in vivo.
214517 - Genomic rna of the murine coronavirus jhm.
427117 - Metabolism of low molecular weight ribonucleic acids in early sea urchin embryos.
10465767 - A molecular model for reca-promoted strand exchange via parallel triple-stranded helices.
22296597 - Recql4 localizes to mitochondria and preserves mitochondrial dna integrity.
Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Virology     Volume:  293     ISSN:  0042-6822     ISO Abbreviation:  Virology     Publication Date:  2002 Feb 
Date Detail:
Created Date:  2002-03-11     Completed Date:  2002-05-01     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0110674     Medline TA:  Virology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  368-78     Citation Subset:  IM    
AIDS Vaccine Program, National Cancer Institute-Frederick, National Institutes of Health, Frederick, Maryland 21702-1201, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Cell Line
Gene Products, gag / genetics,  metabolism,  physiology*
HIV-1 / genetics,  physiology*
Point Mutation
Protein Biosynthesis
Protein Precursors / genetics,  metabolism,  physiology*
RNA, Messenger / metabolism
RNA, Viral / genetics*,  metabolism
Virus Assembly*
Grant Support
Reg. No./Substance:
0/Gene Products, gag; 0/Protein Precursors; 0/RNA, Messenger; 0/RNA, Viral; 0/p55 gag precursor protein, Human immunodeficiency virus 1

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  Equine herpesvirus type 1 devoid of gM and gp2 is severely impaired in virus egress but not direct c...
Next Document:  Evidence of nucleotidyl phosphatase activity associated with core protein sigma A of avian reovirus ...