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PMID: 22496887 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Functional and structural analysis of the internal ribosome entry site present in the mRNA of natural variants of the HIV-1.

PloS one ·Vol. 7 ·No. 4 ·2012-00-00 ·Pages e35031

Vallejos M, Carvajal F, Pino K, Navarrete C, Ferres M, Huidobro-Toro JP, Sargueil B, López-Lastra M

Abstract

The 5'untranslated regions (UTR) of the full length mRNA of the HIV-1 proviral clones pNL4.3 and pLAI, harbor an internal ribosomal entry site (IRES). In this study we extend this finding by demonstrating that the mRNA 5'UTRs of natural variants of HIV-1 also exhibit IRES-activity. Cap-independent translational activity was demonstrated using bicistronic mRNAs in HeLa cells and in Xenopus laevis oocytes. The possibility that expression of the downstream cistron in these constructs was due to alternative splicing or to cryptic promoter activity was ruled out. The HIV-1 variants exhibited significant 5'UTR nucleotide diversity with respect to the control sequence recovered from pNL4.3. Interestingly, translational activity from the 5'UTR of some of the HIV-1 variants was enhanced relative to that observed for the 5'UTR of pNL4.3. In an attempt to explain these findings we probed the secondary structure of the variant HIV-1 5'UTRs using enzymatic and chemical approaches. Yet subsequent structural analyses did not reveal significant variations when compared to the pNL4.3-5'UTR. Thus, the increased IRES-activity observed for some of the HIV-1 variants cannot be ascribed to a specific structural modification. A model to explain these findings is proposed.

MeSH Terms
5' Untranslated Regions HIV-1/metabolism HeLa Cells Humans Models, Biological Nucleic Acid Conformation Protein Biosynthesis RNA, Messenger/metabolism RNA, Viral/blood Ribosomes/metabolism
Chemicals
5' Untranslated Regions RNA, Messenger RNA, Viral
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Vallejos Maricarmen
Laboratorio de Virología Molecular, Instituto Milenio de Inmunología e Inmunoterapia, Centro de Investigaciones Médicas, Escuela de Medicina, Pontificia Universidad Católica de Chile, Santiago, Chile.
Carvajal Felipe
Pino Karla
Navarrete Camilo
Ferres Marcela
Huidobro-Toro Juan Pablo
Sargueil Bruno
López-Lastra Marcelo
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Article Info
Journal
PloS one
Abbr.
PLoS One
ISSN
1932-6203
DOI
10.1371/journal.pone.0035031
Published
2012-00-00
Epub
2012-00-04
Pages
e35031
Language
English
Region
United States
NLM ID
101285081
PMCID
PMC3319624
Subset
IM
External Links
PubMed source DOI: 10.1371/journal.pone.0035031 PMC Full Text View journal details
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