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Volume 270, Number 12, Issue of March 24, 1995 pp. 6991-6996
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Regulation of Saccharomyces cerevisiae Flavohemoglobin Gene Expression (*)

(Received for publication, November 1, 1994; and in revised form, December 27, 1994)

Michael J. Crawford ,  David R. Sherman (§),  ,  Daniel E. Goldberg (¶)

From the Howard Hughes Medical Institute, Departments of Medicine and Molecular Microbiology, Washington University Medical School and the Jewish Hospital of St. Louis, St. Louis, Missouri 63110

ABSTRACT

The Saccharomyces cerevisiae hemoglobin is a flavoprotein of unknown function. It shares extensive sequence homology with the globin of Candida as well as those of several bacterial species. We have studied its gene regulation in order to better understand its purpose in the cell. Transcriptional analyses indicate that, in sharp contrast to the bacterial globins of Vitreoscilla and Alcaligenes eutrophus, the S. cerevisiae globin message is induced during logarithmic growth and under oxygen-replete conditions. Transcription of the S. cerevisiae hemoglobin gene is positively regulated by the transcription factors heme-activated protein (HAP) 1 and HAP2/3/4, which respond to intracellular heme levels. Anaerobically, there is a low level, HAP-independent induction of hemoglobin mRNA. Unlike other systems influenced by the HAP2/3/4 transcription factor complex, no activation of hemoglobin expression by growth in non-fermentable carbon sources is observed. Flavohemoglobin gene disruption does not alter cell viability or growth in a variety of oxygen conditions and carbon sources. Physical and genetic mapping of the S. cerevisiae flavohemoglobin gene places it on chromosome seven near the formyltetrahydrofolate synthase (ADE3) locus. These data indicate that, despite the high degree of homology, the S. cerevisiae globin may have a function distinct from those proposed for bacterial globins.

FOOTNOTES

*
This work was supported in part by a grant from the Lucille P. Markey Charitable Trust. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked ``advertisement'' in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§
Current address: PathoGenesis Corp., 201 Elliott Ave. W., Seattle, WA 98119.

To whom correspondence should be addressed: Dept. of Molecular Microbiology, Washington University School of Medicine, Box 8230, 660 S. Euclid Ave., St. Louis, MO 63110. Tel: 314-362-1514; Fax: 314-362-1232.

(^1)
The abbreviations used are: VtHb, Vitreoscilla hemoglobin; HAP, heme-activated protein; YHG, S. cerevisiae flavohemoglobin; TIF51, translation initiation factor 5; COX5, cytochrome oxidase subunit 5; ACT1, actin; kb, kilobase pair(s); PCR, polymerase chain reaction.

ACKNOWLEDGEMENTS

We would like to thank John Cooper, Leonard Guarente, and Richard Zitomer for the provision of strains and John Lawrence for use of the densitometer. We are indebted to Mark Johnston, Linda Riles, and Tina Hesman for the provision of yeast strains as well as for invaluable discussion.

©1995 by The American Society for Biochemistry and Molecular Biology, Inc.

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