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Molecular and Cellular Biology, March 1999, p. 2044-2050, Vol. 19, No. 3
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Nrg1 Is a Transcriptional Repressor for Glucose Repression of STA1 Gene Expression in Saccharomyces cerevisiae

Seok Hee Park, Sang Seok Koh,dagger Jae Hwan Chun, Hye Jin Hwang, and Hyen Sam Kang*

Department of Microbiology, College of Natural Sciences, Seoul National University, Seoul 151-742, Korea

Received 6 July 1998/Returned for modification 20 August 1998/Accepted 25 November 1998

Expression of genes encoding starch-degrading enzymes is regulated by glucose repression in the yeast Saccharomyces cerevisiae. We have identified a transcriptional repressor, Nrg1, in a genetic screen designed to reveal negative factors involved in the expression of STA1, which encodes a glucoamylase. The NRG1 gene encodes a 25-kDa C2H2 zinc finger protein which specifically binds to two regions in the upstream activation sequence of the STA1 gene, as judged by gel retardation and DNase I footprinting analyses. Disruption of the NRG1 gene causes a fivefold increase in the level of the STA1 transcript in the presence of glucose. The expression of NRG1 itself is inhibited in the absence of glucose. DNA-bound LexA-Nrg1 represses transcription of a target gene 10.7-fold in a glucose-dependent manner, and this repression is abolished in both ssn6 and tup1 mutants. Two-hybrid and glutathione S-transferase pull-down experiments show an interaction of Nrg1 with Ssn6 both in vivo and in vitro. These findings indicate that Nrg1 acts as a DNA-binding repressor and mediates glucose repression of the STA1 gene expression by recruiting the Ssn6-Tup1 complex.

* Corresponding author. Mailing address: Department of Microbiology, College of Natural Sciences, Seoul National University, Shillim-Dong, Kwanak-Gu, Seoul 151-742, Korea. Phone: 82-2-880-6701. Fax: 82-2-876-4440 or 82-2-888-4911. E-mail: khslab{at}khslab.snu.ac.kr.

dagger Present address: Whitehead Institute for Biomedical Research, Cambridge, MA 02142.

Molecular and Cellular Biology, March 1999, p. 2044-2050, Vol. 19, No. 3
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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