SAP1, A PROTEIN THAT BINDS TO SEQUENCES-REQUIRED FOR MATING-TYPE SWITCHING, IS ESSENTIAL FOR VIABILITY IN SCHIZOSACCHAROMYCES-POMBE

The pattern of mating-type switching in cell pedigrees of the fission yeast Schizosaccharomyces pombe is dictated by the inheritance of spec ific DNA chains at the mating-type locus (mat1). The recombination eve nt essential for switching is initiated by a site-specific double-stra nd break at mat1. The switch-activating protein, Sap1, binds in vitro to a mat1 cis-acting site that was shown earlier to be essential for e fficient mating-type switching. We isolated the sap1 gene by using oli gonucleotides corresponding to the amino acid sequence of purified Sap 1 protein. The sequence of that gene predicted a 30-kDa protein with n o significant homology to other canonical DNA-binding protein motifs. To facilitate its biochemical characterization, Sap1 was expressed in Escherichia coli. The protein expressed in bacteria displayed the same DNA-binding specificities as the protein purified from S. pombe. Inte restingly, analysis of a sap1 null mutation showed that the gene is es sential for growth even in a strain in which mating-type snitching is prohibited because of a defect in generation of the double-strand brea k. Thus, the sap1 gene product implicated in mating-type switching is shown to be essential for cell viability.