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. 1989 Feb;121(2):223–236. doi: 10.1093/genetics/121.2.223

The Mfα1 Gene of Saccharomyces Cerevisiae: Genetic Mapping and Mutational Analysis of Promoter Elements

M C Flessel 1, A J Brake 1, J Thorner 1
PMCID: PMC1203612  PMID: 2659433

Abstract

The activity and cell-type specificity of the promoter of the MFα1 gene of Saccharomyces cerevisiae were examined by measuring expression of an MFα1-SUC2 gene fusion in MATa, MATα, and MATa/MATα cells. A high level of invertase activity was observed only in MATα cells. Weak expression occurred in MATa cells when the hybrid gene was carried on a multicopy plasmid or on a centromere-containing plasmid, but not when the hybrid gene was integrated at the normal MFα1 locus. Analysis of a set of 5'-deletions of the promoter region of the MFα1-SUC2 gene on the multicopy plasmid indicated that sequences from -354 to -274 upstream of the translational start site were required for high level expression in MATα cells. Smaller internal deletions and insertions within the promoter region of the MFα1-SUC2 gene were inserted into the genome at the normal MFα1 locus. These mutations further delineated four promoter domains important for expression: (1) two 26 bp elements (-365 to -340 and -312 to -287) with imperfect dyad symmetry; (2) a 40 bp segment (-264 to -226) that lies about 120 bp upstream of the TATA box; and (3) the TATA box itself (-128 to -122). The transcriptional start sites of the normal MFα1 promoter and of a mutant lacking the TATA box were determined. The MFα1 locus was mapped to the left arm of chromosome XVI, about 22 cM centromere-proximal to the PEP4 gene.

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Selected References

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