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Originally published In Press as doi:10.1074/jbc.M009546200 on January 23, 2001

J. Biol. Chem., Vol. 276, Issue 17, 14374-14384, April 27, 2001
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Identification and Characterization of a Low Oxygen Response Element Involved in the Hypoxic Induction of a Family of Saccharomyces cerevisiae Genes
IMPLICATIONS FOR THE CONSERVATION OF OXYGEN SENSING IN EUKARYOTES*

Michael J. VasconcellesDagger §, Yide JiangDagger §, Kevin McDaidDagger , Laura GiloolyDagger , Sharon WretzelDagger , David L. PorterDagger ||, Charles E. Martin**, and Mark A. GoldbergDagger Dagger Dagger

From the Dagger  Hematology Division, Department of Medicine, Brigham & Women's Hospital, and Harvard Medical School, Boston, Massachusetts 02115 and ** The Nelson Biological Laboratory, Bureau of Biological Research, Department of Cell Biology and Neuroscience, Rutgers University, Busch Campus, Piscataway, New Jersey 08854

An organism's ability to respond to changes in oxygen tension depends in large part on alterations in gene expression. The oxygen sensing and signaling mechanisms in eukaryotic cells are not fully understood. To further define these processes, we have studied the Delta 9 fatty acid desaturase gene OLE1 in Saccharomyces cerevisiae. We have confirmed previous data showing that the expression of OLE1 mRNA is increased in hypoxia and in the presence of certain transition metals. OLE1 expression was also increased in the presence of the iron chelator 1,10-phenanthroline. A 142-base pair (bp) region 3' to the previously identified fatty acid response element was identified as critical for the induction of OLE1 in response to these stimuli using OLE1 promoter-lacZ reporter constructs. Electromobility shift assays confirmed the presence of an inducible band shift in response to hypoxia and cobalt. Mutational analysis defined the nonameric sequence ACTCAACAA as necessary for transactivation. A 20-base pair oligonucleotide containing this nonamer confers up-regulation by hypoxia and inhibition by unsaturated fatty acids when placed upstream of a heterologous promoter in a lacZ reporter construct. Additional yeast genes were identified which respond to hypoxia and cobalt in a manner similar to OLE1. A number of mammalian genes are also up-regulated by hypoxia, cobalt, nickel, and iron chelators. Hence, the identification of a family of yeast genes regulated in a similar manner has implications for understanding oxygen sensing and signaling in eukaryotes.

* This work was supported by National Institues of Health Grants DK45098 (to M. A. G.), GM45768 (to C. E. M.), and K08 HL03599 (to M. J. V.).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.

§ Contributed equally to the results of this work.

Present address: Dept. of Adult Oncology, Dana-Farber Cancer Institute, Dept. of Medicine, Brigham and Women's Hospital, and Harvard Medical School, Boston, MA 02115.

|| Present address: Hematology-Oncology Div., University of Pennsylvania Medical Center, Philadelphia, PA 19104.

Dagger Dagger To whom correspondence should be addressed: Hematology Div., Dept. of Medicine, Brigham & Women's Hospital, Harvard Medical School, 221 Longwood Ave., Boston, MA 02115. Tel.: 617-732-5841; Fax: 617-739-0748; E-mail: Mark.Goldberg@genzyme.com.

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