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M9470036.TXT
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1994-07-02
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Document 0036
DOCN M9470036
TI Identification of the primary site of the human immunodeficiency virus
type 1 RNA dimerization in vitro.
DT 9409
AU Skripkin E; Paillart JC; Marquet R; Ehresmann B; Ehresmann C; Unite
Propre de Recherche 9002 du Centre National de la; Recherche
Scientifique, Institut de Biologie Moleculaire et; Cellulaire,
Strasbourg, France.
SO Proc Natl Acad Sci U S A. 1994 May 24;91(11):4945-9. Unique Identifier :
AIDSLINE MED/94255445
AB The diploid genome of all retroviruses is made of two homologous copies
of RNA intimately associated near their 5' end, in a region called the
dimer linkage structure. Dimerization of genomic RNA is thought to be
important for crucial functions of the retroviral life cycle (reverse
transcription, translation, encapsidation). Previous in vitro studies
mapped the dimer linkage structure of human immunodeficiency virus type
1 (HIV-1) in a region downstream of the splice donor site, containing
conserved purine tracts that were postulated to mediate dimerization,
through purine quartets. However, we recently showed that dimerization
of HIV-1 RNA also involves sequences upstream of the splice donor site.
Here, we used chemical modification interference to identify nucleotides
that are required in unmodified form for dimerization of a RNA fragment
containing nucleotides 1-707 of HIV-1 RNA. These nucleotides map
exclusively in a restricted area upstream of the splice donor site and
downstream of the primer binding site. They are centered around a
palindromic sequence (GUGCAC279) located in a hairpin loop. Our results
support a model in which dimer formation is initiated by the annealing
of the palindromic sequences, possibly by a loop-loop interaction
between the two monomers. Further experiments show that the deletion of
the stem-loop or base substitutions in the loop abolish dimerization,
despite the presence of the previously postulated dimer linkage
structure. On the other hand, deletions of the purine tracts downstream
of the splice donor site do not prevent dimerization. Therefore, we
conclude that the palindromic region represents the dimerization
initiation site of genomic RNA.
DE Base Sequence Binding Sites Biopolymers HIV-1/*GENETICS Molecular
Sequence Data Nucleic Acid Conformation Nucleotides/CHEMISTRY
Repetitive Sequences, Nucleic Acid RNA, Viral/*CHEMISTRY Support,
Non-U.S. Gov't JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).