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M94A3299.TXT
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1994-10-25
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Document 3299
DOCN M94A3299
TI In vivo interaction of HIV-1 Rev with an extended Stem I duplex
structure of Rev response element RNA.
DT 9412
AU Kimura T; Department of Microbiology, Kansai Medical University, Osaka,;
Japan.
SO Int Conf AIDS. 1994 Aug 7-12;10(1):108 (abstract no. PA0049). Unique
Identifier : AIDSLINE ICA10/94369276
AB OBJECTIVE: Experiments were designed to study whether the packaging
reaction of HIV-1 Rev with Rev response element (RRE) RNA reported in
vitro mediates Rev function in vivo. METHODS: RRE activity was examined
using HIV-1-derived reporter plasmids, which contained the bacterial
chloramphenicol acetyltransferase (CAT) gene as a reporter. RESULTS: The
originally defined 234-residue RRE failed to show the full Rev response
when inserted in a test plasmid that lacks the flanking env sequences. A
computer search for additional RNA secondary structures in the flanking
regions revealed an alternative 351-residue complete RRE, which carried
an extended Stem I duplex RNA structure. Mutations in the complete RRE
that disrupted either the primary Rev-binding bubble within Stem-loop II
or the duplex RNA structure in the extended Stem I region showed reduced
CAT activities with various degrees. DISCUSSION AND CONCLUSIONS: The
mutagenesis studies showed that the 351-residue complete RRE rather than
the original 234-residue RRE was required to exert the full
Rev-responsiveness in vivo. For the full responsiveness, it was found
that association of Rev with RRE was to be initiated with the
recognition of the functional bubble structure in the Stem-loop II,
followed by the interaction with the neighbouring duplex RNA, including
the elongated Stem I structure.
DE Binding Sites/GENETICS Chloramphenicol Acetyltransferase/GENETICS Gene
Products, rev/CHEMISTRY/*GENETICS/*METABOLISM Genes, env Genes,
Reporter Human HIV-1/*GENETICS/*METABOLISM In Vitro Mutagenesis
Nucleic Acid Conformation Plasmids/GENETICS RNA,
Viral/CHEMISTRY/GENETICS/*METABOLISM MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).