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Originally published In Press as doi:10.1074/jbc.M101233200 on April 17, 2001

J. Biol. Chem., Vol. 276, Issue 27, 25279-25286, July 6, 2001
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Pdx1 Level Defines Pancreatic Gene Expression Pattern and Cell Lineage Differentiation*

Haiyan WangDagger §, Pierre MaechlerDagger , Beate Ritz-Laser, Kerstin A. HagenfeldtDagger , Hisamitsu IshiharaDagger , Jacques Philippe, and Claes B. WollheimDagger §

From the Dagger  Division of Clinical Biochemistry and the  Diabetes Unit, Department of Internal Medicine, Geneva University Medical Center, CH-1211 Geneva 4, Switzerland

The absence of Pdx1 and the expression of brain-4 distinguish alpha -cells from other pancreatic endocrine cell lineages. To define the transcription factor responsible for pancreatic cell differentiation, we employed the reverse tetracycline-dependent transactivator system in INS-I cell-derived subclones INSralpha beta and INSrbeta to achieve tightly controlled and conditional expression of wild type Pdx1 or its dominant-negative mutant, as well as brain-4. INSralpha beta cells express not only insulin but also glucagon and brain-4, while INSrbeta cells express only insulin. Overexpression of Pdx1 eliminated glucagon mRNA and protein in INSralpha beta cells and promoted the expression of beta -cell-specific genes in INSrbeta cells. Induction of dominant-negative Pdx1 in INSralpha beta cells resulted in differentiation of insulin-producing beta -cells into glucagon-containing alpha -cells without altering brain4 expression. Loss of Pdx1 function alone in INSrbeta cells, which do not express endogenous brain-4 and glucagon, was also sufficient to abolish the expression of genes restricted to beta -cells and to cause alpha -cell differentiation. In contrast, induction of brain-4 in INSrbeta cells initiated detectable expression of glucagon but did not affect beta -cell-specific gene expression. In conclusion, Pdx1 confers the expression of pancreatic beta -cell-specific genes, such as genes encoding insulin, islet amyloid polypeptide, Glut2, and Nkx6.1. Pdx1 defines pancreatic cell lineage differentiation. Loss of Pdx1 function rather than expression of brain4 is a prerequisite for alpha -cell differentiation.

* This work was supported by Swiss National Science Foundation Grant Number 32-49755.96, the Institute for Human Genetics and Biochemistry, the Berger Foundation, the Carlos and Elsie de Reuters Foundation, and the Juvenile Diabetes Foundation International, New York.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 may be addressed: Division de Biochimie Clinique et de Diabétologie Expérimentale, Départment de Médecine Interne, Center Médical Universitaire, CH-1211 Geneva 4, Switzerland. Tel.: 41 22 702 5548; Fax: 41 22 702 5543; E-mail: Haiyan. Wang@medicine.unige.ch or Claes.Wollheim@medicine.unige.ch.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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