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M9470048.TXT
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1994-07-02
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Document 0048
DOCN M9470048
TI Presence of negative and positive cis-acting RNA splicing elements
within and flanking the first tat coding exon of human immunodeficiency
virus type 1.
DT 9409
AU Amendt BA; Hesslein D; Chang LJ; Stoltzfus CM; Department of
Microbiology, University of Iowa, Iowa City 52242.
SO Mol Cell Biol. 1994 Jun;14(6):3960-70. Unique Identifier : AIDSLINE
MED/94254853
AB The human immunodeficiency virus type 1 (HIV-1) RNA follows a complex
splicing pathway in which a single primary transcript either remains
unspliced or is alternatively spliced to more than 30 different singly
and multiply spliced mRNAs. We have used an in vitro splicing assay to
identify cis elements within the viral genome that regulate HIV-1 RNA
splicing. A novel splicing regulatory element (SRE) within the first tat
coding exon has been detected. This element specifically inhibits
splicing at the upstream 3' splice site flanking this tat exon. The
element only functions when in the sense orientation and is position
dependent when inserted downstream of a heterologous 3' splice site. In
vivo, an HIV-1 SRE mutant demonstrated a decrease in unspliced viral
RNA, increased levels of single- and double-spliced tat mRNA, and
reduced levels of env and rev mRNAs. In addition to the negative
cis-acting SRE, the flanking 5' splice site downstream of the first tat
coding exon acts positively to increase splicing at the upstream 3'
splice sites. These results are consistent with hypotheses of bridging
interactions between cellular factors that bind to the 5' splice site
and those that bind at the upstream 3' splice site.
DE *Alternative Splicing Base Sequence Cell Nucleus/METABOLISM DNA
Primers *Exons Gene Expression Regulation, Viral *Genes, tat Hela
Cells Human HIV-1/*GENETICS/METABOLISM Molecular Sequence Data
Mutagenesis Polymerase Chain Reaction Regulatory Sequences, Nucleic
Acid Restriction Mapping *RNA Splicing RNA, Messenger/*BIOSYNTHESIS
Support, Non-U.S. Gov't Support, U.S. Gov't, P.H.S. Transfection
JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).