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M9610320.TXT
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1996-01-30
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Document 0320
DOCN M9610320
TI Formation of stable and functional HIV-1 nucleoprotein complexes in
vitro.
DT 9601
AU Tanchou V; Gabus C; Rogemond V; Darlix JL; LaboRetro, Unite de Virologie
humaine (INSERM-ENS 412), Ecole; Normale Superieure, Lyon, France.
SO J Mol Biol. 1995 Oct 6;252(5):563-71. Unique Identifier : AIDSLINE
MED/96017727
AB HIV genomic RNA resides within the nucleocapsid, in the interior of the
virus, which serves to protect the RNA against nuclease degradation and
to promote its reverse transcription. To investigate the role of
nucleocapsid protein (NCp7) in the stability and replication of genomic
RNA within the nucleocapsid, we used NCp7, reverse transcriptase (RT)
and RNAs representing the 5' and 3' regions of the genome to
reconstitute functional HIV-1 nucleocapsids. The nucleoprotein complexes
generated in vitro were found to be stable, which, according to
biochemical and genetic data, probably results from the tight binding of
NCp7 molecules to the RNA and strong NCp7/NCp7 interactions. The
nucleoprotein complexes efficiently protected viral RNA against RNase
degradation and, at the same time, promoted viral DNA synthesis by RT.
DNA strand transfer from the 5' to the 3' RNA template was very
efficient in nucleoprotein complexes formed in the presence of both
RNAs, but not when the RNAs were in separate complexes. These results
indicate that the in vitro reconstituted HIV-1 nucleoprotein complexes
function like virion nucleocapsids and thus provide a way to study at
the molecular level this viral substructure and the synthesis of
proviral DNA, and to search for new anti-HIV agents.
DE Base Sequence Capsid/GENETICS/*METABOLISM DNA Primers DNA,
Complementary/BIOSYNTHESIS DNA, Single-Stranded/BIOSYNTHESIS DNA,
Viral/*BIOSYNTHESIS Electrophoresis, Polyacrylamide Gel Gene Products,
gag/GENETICS/*METABOLISM Human HIV-1/GENETICS/*METABOLISM Molecular
Sequence Data Polymerase Chain Reaction Recombinant Fusion
Proteins/GENETICS Ribonuclease T1/METABOLISM RNA-Directed DNA
Polymerase/METABOLISM RNA, Viral/GENETICS/*METABOLISM Support,
Non-U.S. Gov't Templates Zinc Fingers JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).