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J Biol Chem, Vol. 275, Issue 19, 14155-14166, May 12, 2000

Glucose-regulated Turnover of mRNA and the Influence of Poly(A) Tail Length on Half-life^*

Susana Prieto, Bernard J. de la Cruz^§, and Immo E. Scheffler^¶

From the Department of Biology, University of California, La Jolla, California 92093-0322

Glucose repression in Saccharomyces cerevisiae can now be seen to operate at two levels: regulation of transcription of certain genes and control of the half-life of the corresponding mRNAs (Scheffler, I. E., de la Cruz, B. J., and Prieto, S. (1998) Int. J. Biochem Cell Biol. 30, 1175-1193). For example, the steady state levels of SDH2 mRNA and SUC2 mRNA are significantly determined by their differential rates of turnover. A current model for the mechanism of mRNA turnover includes three distinct steps: a rate-limiting deadenylation, removal of the 5'-7-methyl-G (decapping), and 5'-3' exonuclease digestion. We have investigated the same three reactions during glucose-induced degradation of these transcripts. Our results indicate that while decapping (by Dcp1p) and 5'-3' exonuclease digestion (by Xrn1p) are obligatory steps for the rapid degradation of these mRNAs, the dependence on deadenylation is more complicated. At steady state in glycerol these transcripts have very short poly(A) tails but are nevertheless very stable; the addition of glucose causes immediate decapping and degradation without further deadenylation; in contrast, newly made SUC2 mRNA (after a shift from glucose to glycerol) has significantly longer poly(A) tails, and such transcripts are not rapidly degraded upon addition of glucose. A constitutive deadenylation reaction that is independent of the carbon source eventually makes the stability of these transcripts very sensitive to glucose. These results are interpreted in terms of a working hypothesis proposing a competition between translational initiation and decapping influenced by the carbon source. The presence of a long poly(A) tail may also affect this competition in favor of translational initiation and mRNA stabilization.

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^* This work was supported in part by grants from the National Science Foundation and the American Cancer Society (to I. E. S.).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.

Postdoctoral fellow supported by the Spanish Ministerio de Educacion y Cultura.

^§ Supported by a training grant from the United States Public Health Service.

^¶ To whom correspondence should be addressed. Tel.: 858-534-2741; Fax: 858-534-0053; E-mail: ischeffler@ucsd.edu.

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Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.

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