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Molecular and Cellular Biology, September 2001, p. 6161-6169, Vol. 21, No. 18
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.18.6161-6169.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

MGA2 Is Involved in the Low-Oxygen Response Element-Dependent Hypoxic Induction of Genes in Saccharomyces cerevisiae

Yide Jiang,1 Michael J. Vasconcelles,1,dagger Sharon Wretzel,1 Anne Light,1 Charles E. Martin,2 and Mark A. Goldberg1,*

Hematology Division, Department of Medicine, Brigham & Women's Hospital, and Harvard Medical School, Boston, Massachusetts 02115,1 and Nelson Biological Laboratory, Bureau of Biological Research, Department of Cell Biology and Neuroscience, Rutgers University, Busch Campus, Piscataway, New Jersey 088542

Received 3 May 2001/Returned for modification 30 May 2001/Accepted 12 June 2001

Eukaryotes have the ability to respond to changes in oxygen tension by alterations in gene expression. For example, OLE1 expression in Saccharomyces cerevisiae is upregulated under hypoxic conditions. Previous studies have suggested that the pathway regulating OLE1 expression by unsaturated fatty acids may involve Mga2p and Spt23p, two structurally and functionally related proteins. To define the possible roles of each of these genes on hypoxia-induced OLE1 expression, we examined OLE1 expression under normoxia, hypoxia, and cobalt treatment conditions in Delta mga2 or Delta spt23 deletion strains. The results of OLE1 promoter-lacZ reporter gene and Northern blot analyses showed that hypoxia- and cobalt-induced OLE1 expression was dramatically decreased in a Delta mga2 strain but not in a Delta spt23 strain. Further analyses using low-oxygen response element (LORE)-CYC1-lacZ fusion reporter assays and electrophoretic mobility shift assays (EMSAs) demonstrated that MGA2 significantly affects the LORE-dependent hypoxic induction pathway of gene expression. When MGA2 was supplied by a plasmid, the LORE-dependent hypoxia-inducible reporter expression was recovered, as was the hypoxia-inducible complex in EMSAs in the S. cerevisiae Delta mga2 strain. Supershift analysis of EMSAs using crude extracts containing mycMga2p indicated that Mga2p is a component of the LORE-binding complex. Another LORE-dependent, hypoxia-inducible gene, ATF1, was similarly affected in the Delta mga2 strain. These results indicate that MGA2 is required for the LORE-dependent hypoxic gene induction in S. cerevisiae.

* Corresponding author. Mailing address: Hematology Division, Department of Medicine, Brigham & Women's Hospital, Harvard Medical School, 221 Longwood Ave., Boston, MA 02115. Phone: (617) 732-5841. Fax: (617) 739-0748. E-mail: Mark.Goldberg{at}genzyme.com.

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

Molecular and Cellular Biology, September 2001, p. 6161-6169, Vol. 21, No. 18
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.18.6161-6169.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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