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J. Biol. Chem., Vol. 276, Issue 13, 10514-10523, March 30, 2001
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From the Department of Biochemistry, West Virginia University,
Morgantown, West Virginia 26506
Expression of glucose-6-phosphate dehydrogenase
(G6PD) gene during starvation and refeeding is regulated by a
posttranscriptional mechanism occurring in the nucleus. The amount of
G6PD mRNA at different stages of processing was measured in RNA
isolated from the nuclear matrix fraction of mouse liver. This nuclear
fraction contains nascent transcripts and RNA undergoing processing.
Using a ribonuclease protection assay with probes that cross an
exon-intron boundary in the G6PD transcript, the abundance of mRNAs
that contain the intron (unspliced) and without the intron (spliced)
was measured. Refeeding resulted in 6- and 8-fold increases in
abundance of G6PD unspliced and spliced RNA, respectively, in the
nuclear matrix fraction. However, the amount of G6PD unspliced RNA was
at most 15% of the amount of spliced RNA. During refeeding, G6PD
spliced RNA accumulated at a rate significantly greater than unspliced RNA. Further, the amount of partially spliced RNA exceeded the amount
of unspliced RNA indicating that the enhanced accumulation occurs early
in processing. Starvation and refeeding did not regulate either the
rate of polyadenylation or the length of the poly(A) tail. Thus, the
G6PD gene is regulated during refeeding by enhanced efficiency of
splicing of its RNA, and this processing protects the mRNA from
decay, a novel mechanism for nutritional regulation of gene expression.
Regulation of the Processing of Glucose-6-phosphate
Dehydrogenase mRNA by Nutritional Status*
*
This work was supported by Grant DK46897 from the National
Institutes of Health.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Biochemistry,
West Virginia University, Health Sciences Center, P. O. Box 9142, Morgantown, WV 26506. Tel.: 304-293-7759; Fax: 304-293-6846; E-mail: lsalati@hsc.wvu.edu.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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