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Originally published In Press as doi:10.1074/jbc.M003335200 on July 25, 2000

J. Biol. Chem., Vol. 275, Issue 40, 31069-31077, October 6, 2000
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Sterol Regulatory Element-binding Protein-1 Is Regulated by Glucose at the Transcriptional Level*

Alyssa H. HastyDagger §, Hitoshi Shimano||, Naoya YahagiDagger , Michiyo Amemiya-KudoDagger , Stéphane PerreyDagger , Tomohiro YoshikawaDagger , Jun-ichi OsugaDagger , Hiroaki OkazakiDagger , Yoshiaki TamuraDagger , Yoko IizukaDagger , Futoshi ShionoiriDagger , Ken OhashiDagger , Kenji HaradaDagger , Takanari GotodaDagger , Ryozo NagaiDagger , Shun IshibashiDagger , and Nobuhiro Yamada

From the Dagger  Department of Metabolic Diseasese, University of Tokyo, Tokyo 113-8655, Japan and the  Department of Internal Medicine, Institute of Clinical Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan

In vivo studies suggest that sterol regulatory element-binding protein (SREBP)-1 plays a key role in the up-regulation of lipogenic genes in the livers of animals that have consumed excess amounts of carbohydrates. In light of this, we sought to use an established mouse hepatocyte cell line, H2-35, to further define the mechanism by which glucose regulates nuclear SREBP-1 levels. First, we show that these cells transcribe high levels of SREBP-1c that are increased 4-fold upon differentiation from a prehepatocyte to a hepatocyte phenotype, making them an ideal cell culture model for the study of SREBP-1c induction. Second, we demonstrate that the presence of precursor and mature forms of SREBP-1 protein are positively regulated by medium glucose concentrations ranging from 5.5 to 25 mM and are also regulated by insulin, with the amount of insulin in the fetal bovine serum being sufficient for maximal stimulation of SREBP-1 expression. Third, we show that the increase in SREBP-1 protein is due to an increase in SREBP-1 mRNA. Reporter gene analysis of the SREBP-1c promoter demonstrated a glucose-dependent induction of transcription. In contrast, expression of a fixed amount of the precursor form of SREBP-1c protein showed that glucose does not influence its cleavage. Fourth, we demonstrate that the glucose induction of SREBP could not be reproduced by fructose, xylose, or galactose nor by glucose analogs 2-deoxy glucose and 3-O-methyl glucopyranose. These data provide strong evidence for the induction of SREBP-1c mRNA by glucose leading to increased mature protein in the nucleus, thus providing a potential mechanism for the up-regulation of lipogenic genes by glucose in vivo.

* This work was supported in part by Promotion of Fundamental Studies in Health Science of the Organization for Pharmaceutical Safety and Research and Health Sciences Research Grants (Research on Human Genome and Gene Therapy) from the Ministry of Health and Welfare.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.

§ Recipient of a fellowship under the Japan Society for the Promotion of Science Postdoctoral Fellowship Program for Foreign Researchers.

|| To whom correspondence should be addressed: Dept. of Internal Medicine, Institute of Clinical Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan. Fax: 81-298-53-3053; E-mail: shimano-tky@umin.ac.jp.

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