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).