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Journal of Bacteriology, August 1998, p. 4177-4183, Vol. 180, No. 16
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

A Mutation in a Purported Regulatory Gene Affects Control of Sterol Uptake in Saccharomyces cerevisiae

James H. Crowley,dagger Frank W. Leak Jr., Kevin V. Shianna, Shirley Tove, and Leo W. Parks*

Department of Microbiology, North Carolina State University, Raleigh, North Carolina 27695-7615

Received 11 February 1998/Accepted 14 June 1998

Aerobically growing wild-type strains of Saccharomyces cerevisiae are unable to take exogenously supplied sterols from media. This aerobic sterol exclusion is vitiated under anaerobic conditions, in heme-deficient strains, and under some conditions of impaired sterol synthesis. Mutants which can take up sterols aerobically in heme-competent cells have been selected. One of these mutations, designated upc2-1, gives a pleiotropic phenotype in characteristics as diverse as aerobic accumulation of sterols, total lipid storage, sensitivity to metabolic inhibitors, response to altered sterol structures, and cation requirements. During experiments designed to ascertain the effects of various cations on yeast with sterol alterations, it was observed that upc2-1 was hypersensitive to Ca2+. Using resistance to Ca2+ as a screening vehicle, we cloned UPC2 and showed that it is YDR213W, an open reading frame on chromosome IV. This belongs to a fungal regulatory family containing the Zn(II)2Cys6 binuclear cluster DNA binding domain. The single guanine-to-adenine transition in upc2-1 gives a predicted amino acid change from glycine to aspartic acid. The regulatory defect explains the semidominance and pleiotropic effects of upc2-1.

* Corresponding author. Mailing address: Department of Microbiology, North Carolina State University, Raleigh, NC 27695-7615. Phone: (919) 515-7860. Fax: (919) 515-7867. E-mail: parks{at}mbio.ncsu.edu.

dagger Present address: NutraSweet Kelco Co., San Diego, CA 92123-1718.

Journal of Bacteriology, August 1998, p. 4177-4183, Vol. 180, No. 16
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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