Cloning and analysis of transcription of the mei2 gene responsible for initiation of meiosis in the fission yeast Schizosaccharomyces pombe

Abstract

We have isolated a hybrid plasmid, pDB(mei2)2, containing a 7.4-kilobases (kb) DNA fragment from a Schizosaccharomyces pombe genomic library which is able to complement the mei2 mutation of S. pombe. Integration of the cloned DNA sequence at the mei2 site on chromosome I demonstrated that it contained the mei2 gene. This gene was localized on a 4.7-kb HindIII-PvuII fragment in the subclone pFMV402. Transcriptional regulation was studied by Northern blot analysis in which polyadenylated RNA was prepared from a heterozygous (h+N/h-S) diploid strain cultured either in nitrogen-rich growth medium or in nitrogen-free sporulation medium. The size of the major mei2 mRNA, which always gave a broad band, was estimated to be 4.2 +/- 0.2 kb, and a few minor bands (e.g., 3.2 and 1.8 kb) appeared as well. These transcripts appeared more abundantly in sporulating cells than in growing cells. Neither the mating type genes (mat) nor the mei3 gene was essential for transcription of the mei2 gene, since ample mei2 mRNA was detected in sporulation-deficient cells transferred to sporulation medium, such as h+N/h+N and h-S/h-S homozygotes, as well as mei1 and mei3 mutants.