ABSTRACT
In order to isolate meiosis-specific genes in Schizosaccharomyces pombe, we have constructed a subtracted cDNA library enriched in clones whose expression is enhanced during meiosis induced by nitrogen starvation. Using northern blot analysis, we isolated 31 kinds of clones whose expression was induced in a meiosis/sporulation-specific manner. We comprehensively named them meu after meiotic expression upregulated. The transcription of 20 meu genes was found to be dependent on the mei4(+) gene, which encodes a transcription factor required for the progression of meiosis. DNA sequencing indicated that most of the meu genes encode novel proteins. Notably, five of the meu genes harbor no apparent protein coding sequences, and the transcripts form stable hairpin structures, suggesting that they may generate non-coding RNAs or antisense RNAS: The results presented here imply that RNAs are also important for the comprehensive characterization of genomic expression.
Subject(s)
Fungal Proteins/genetics , Gene Expression Regulation, Fungal , Meiosis/genetics , Schizosaccharomyces/genetics , Base Sequence , DNA, Complementary/genetics , DNA, Complementary/isolation & purification , Gene Library , Genes, Fungal/genetics , Molecular Sequence Data , RNA, Fungal/genetics , Schizosaccharomyces/physiology , Sequence Homology, Nucleic Acid , Spores, Fungal/genetics , Transcription, GeneticABSTRACT
We report here a Schizosaccharomyces pombe gene (dmc1(+)) that resembles budding yeast DMC1 in the region immediately upstream of the rad24(+) gene. We showed by northern and Southern blot analysis that dmc1(+) and rad24(+) are co-transcribed as a bicistronic mRNA of 2.8 kb with meiotic specificity, whereas rad24(+) itself is constitutively transcribed as a 1.0-kb mRNA species during meiosis. Induction of the bicistronic transcript is under the control of a meiosis-specific transcription factor, Ste11. Disruption of both dmc1(+) and rad24(+) had no effect on mitosis or spore formation, and dmc1Delta cells displayed no change in sensitivity to UV or gamma irradiation relative to the wild type. Tetrad analysis indicated that Dmc1 is involved in meiotic recombination. Analysis of gene conversion frequencies using single and double mutants of dmc1 and rhp51 indicated that both Dmc1 and Rhp51 function in meiotic gene conversion. These observations, together with a high level of sequence identity, indicate that the dmc1(+) gene of S. POMBE: is a structural homolog of budding yeast DMC1, sharing both similar and distinct functions in meiosis.
Subject(s)
DNA-Binding Proteins/physiology , Meiosis/genetics , Recombinases/physiology , Recombination, Genetic , Schizosaccharomyces pombe Proteins/physiology , Schizosaccharomyces/genetics , Cell Cycle Proteins/genetics , Cell Division , Crossing Over, Genetic/genetics , DNA, Complementary/genetics , DNA-Binding Proteins/genetics , Fungal Proteins/physiology , Gene Conversion/genetics , Gene Expression Regulation, Fungal , Genes, Fungal/genetics , Intracellular Signaling Peptides and Proteins , Molecular Sequence Data , Mutation , Open Reading Frames/genetics , RNA, Fungal/genetics , RNA, Fungal/isolation & purification , Recombinases/genetics , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae Proteins , Schizosaccharomyces pombe Proteins/genetics , Transcription Factors/physiology , Transcription, GeneticABSTRACT
The rad24(+) gene of Schizosaccharomyces pombe encodes a ubiquitously expressed 14-3-3 protein. We report here that Deltarad24 cells displayed a defect in diploid colony formation, although they conjugated efficiently. We found that a cumulative deletion of mei2(+) gene almost completely suppressed this defect, and demonstrated using two-hybrid analysis that Rad24 protein directly associates with Mei2 protein by recognizing Ser-438 which is a phosphorylation target of Pat1 kinase. We conclude that constitutive progression to meiosis, caused by lack of Mei2 inhibition due to the absence of Rad24 protein, is the primary cause of the proliferative deficiency observed in Deltarad24 cells.
