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(Received for publication, March 11, 1996, and in revised form, August 5, 1996)
From the Cat-1 is a protein with a dual function, a high
affinity, low capacity cationic amino acid transporter of the
y+ system and the receptor for the ecotropic retrovirus. We
have suggested that Cat-1 is required in the regenerating liver for the
transport of cationic amino acids and polyamines in the late G1 phase, a process that is essential for liver cells to
enter mitosis. In our earlier studies we had shown that the
cat-1 gene is silent in the quiescent liver but is induced
in response to hormones, insulin, and glucocorticoids, and partial
hepatectomy. Here we demonstrate that cat-1 is a classic
delayed early growth response gene in the regenerating liver, since
induction of its expression is sensitive to cycloheximide, indicating
that protein synthesis is required. The peak of accumulation of the
cat-1 mRNA (9-fold) by 3 h was not associated with
increased transcriptional activity of the cat-1 gene in the
regenerating liver, indicating post-transcriptional regulation of
expression of this gene. Induction of the cat-1 gene
results in the accumulation of two mRNA species (7.9 and 3.4 kilobase pairs (kb)). Both mRNAs hybridize with the previously
described rat cat-1/2.9-kb cDNA clone. However, the 3
Volume 271, Number 47,
Issue of November 22, 1996
pp. 29799-29806
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
,
,
,
,
Department of Nutrition, Case Western
Reserve University School of Medicine, Cleveland, Ohio 44106 and the
§ Departments of Pediatrics and Cell Biology, Baylor College
of Medicine, Houston, Texas 77030
end of a longer rat cat-1 cDNA (rat
cat-1/6.5-kb) hybridizes only to the 7.9-kb mRNA
transcript. Sequence analysis of this clone indicated that the two
mRNA species result from the use of alternative polyadenylation
signals. The 6.5-kb clone contains a number of AT-rich mRNA
destabilizing sequences which is reflected in the half-life of the
cat-1 mRNAs (90 min for 7.9-kb mRNA and 250 min for
3.4-kb mRNA). Treatment of rats with cycloheximide superinduces the
level of the 7.9-kb cat-1 mRNA in the kidney, spleen,
and brain, but not in the liver, suggesting that cell type-specific
labile factors are involved in its regulation. We conclude that the
need for protein synthesis for induction of the cat-1
mRNA, the short lived nature of the mRNAs, and the multiple sites for regulation of gene expression indicate a tight control of
expression of the cat-1 gene within the regenerating liver and suggest that y+ cationic amino acid transport in liver
cells is regulated at the molecular level.
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