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M9620572.TXT
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1996-02-26
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Document 0572
DOCN M9620572
TI Inefficient spliceosome assembly and abnormal branch site selection in
splicing of an HIV-1 transcript in vitro.
DT 9602
AU Dyhr-Mikkelsen H; Kjems J; Department of Molecular Biology, University
of Aarhus, Denmark.
SO J Biol Chem. 1995 Oct 13;270(41):24060-6. Unique Identifier : AIDSLINE
MED/96025786
AB Continuous replication of human immunodeficiency virus type I (HIV-1)
requires balanced expression of spliced and nonspliced mRNAs in the
cytoplasm. This process is regulated post-transcriptionally by the
viral-encoded Rev protein. An important prerequisite for Rev
responsiveness is the presence of weak splice sites in the viral mRNA.
We have investigated the splicing of the second intron of the HIV-1
Tat/Rev transcript in vitro and show that the 3'-splice site region is
responsible for the inefficient splicing of the HIV-1 transcript. In
contrast, the HIV-1 5'-splice site is highly functional in combination
with a heterologous 3'-splice site. Incubation of the HIV-1 transcript
in nuclear extract leads to a rapid accumulation of 50 S nonproductive
pre-spliceosome complexes. These complexes contain mainly U1 and U2
small nuclear ribonucleoproteins and are formed independently of the
presence of the downstream 3'-splice site. The HIV-1 transcripts, which
do proceed through the first splicing step, utilize primarily a uridine
as the branch acceptor nucleotide. Sequence comparison with other HIV-1
introns suggests that nucleotides other than adenosines are commonly
used as branch points in these viruses.
DE Base Sequence Blotting, Northern DNA Primers Gene Products,
rev/*BIOSYNTHESIS Gene Products, tat/BIOSYNTHESIS Human
HIV-1/*GENETICS/*METABOLISM Molecular Sequence Data Plasmids
Restriction Mapping Ribonucleoproteins, Small Nuclear/METABOLISM RNA
Precursors/*METABOLISM *RNA Splicing RNA, Messenger/*BIOSYNTHESIS
RNA, Small Nuclear/BIOSYNTHESIS Spliceosomes/*METABOLISM Support,
Non-U.S. Gov't *Transcription, Genetic JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).