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M94A3165.TXT
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1994-10-25
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Document 3165
DOCN M94A3165
TI Splicing signals are required for the expression of unspliced
Gag-containing message of moloney murine leukemia virus.
DT 9412
AU Odawara T; Ohshima M; Matano T; Iwamoto A; Yoshikura H; Faculty of
Medicine, University of Tokyo, Japan.
SO Int Conf AIDS. 1994 Aug 7-12;10(1):138 (abstract no. PA0173). Unique
Identifier : AIDSLINE ICA10/94369410
AB OBJECTIVE: All the retroviruses express both unspliced and spliced
messages simultaneously. However, little is known about the control of
the production of the unspliced message under the presence of splicing.
We studied the role of splicing signals on the expression of a
retroviral unspliced message. METHODS: A Moloney murine leukemia virus
(MLV)-derived construct, pG3.6, containing gag as a sole coding region
expressed only small amounts of Gag protein owing to a low message
level. The expression of Gag was restored by the insertion of 2.8k bp
int-env fragment downstream of gag in pG3.6 (pGE6.4). We determined the
sequence in the 2.8k bp fragment required for the expression of the
unspliced message encoding gag. RESULTS: Inversion or deletion of 441 bp
sequence containing a splice acceptor signal in the 2.8k bp int-env
fragment inserted in the downstream of gag abolished the ability of
increasing the gag message, and insertion of the 441 bp alone in the
downstream of gag restored the expression level to that of pGE6.4.
Comparison of nuclear and cytoplasmic RNA levels of pG3.6 with those of
pGE6.4 indicated that the nuclear-cytoplasmic transport of pG3.6 message
was impaired. DISCUSSION AND CONCLUSIONS: For the expression of the
unspliced message encoding gag, MLV genome should be equipped with the
441 bp sequence containing the splice acceptor signal. If the gag gene
of pG3.6 or pGE6.4 was replaced with a neomycin resistance gene, the
expression level was the same for the two constructs. Gag sequence may
contain an inhibitory signal on the expression of RNA lacking the splice
signal.
DE Animal Gene Expression Gene Products, gag/*BIOSYNTHESIS Genes, env
*Genes, gag Genome, Viral Moloney Leukemia Virus/*GENETICS/METABOLISM
Mutagenesis, Insertional RNA Splicing RNA,
Messenger/BIOSYNTHESIS/METABOLISM RNA, Viral/BIOSYNTHESIS/METABOLISM
Signal Peptides/METABOLISM MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).