apparent defect in yeast bud-site selection due to a specific failure to splice the pre-mrna of a regulator of cell-type-specific transcription明显的缺陷在酵母bud-site选择由于特定未能拼接pre-mrna细胞类型特异的转录监管机构的.pdf

Apparent Defect in Yeast Bud-Site Selection Due to a Specific Failure to Splice the Pre-mRNA of a Regulator of Cell-Type-Specific Transcription Shanshan Tuo, Kenichi Nakashima, John R. Pringle* Department of Genetics, Stanford University School of Medicine, Stanford, California, United States of America Abstract The yeast Saccharomyces cerevisiae normally selects bud sites (and hence axes of cell polarization) in one of two distinct patterns, the axial pattern of haploid cells and the bipolar pattern of diploid cells. Although many of the proteins involved in bud-site selection are known, it is likely that others remain to be identified. Confirming a previous report (Ni and Snyder, 2001, Mol. Biol. Cell 12, 2147–2170), we found that diploids homozygous for deletions of IST3/SNU17 or BUD13 do not show normal bipolar budding. However, these abnormalities do not reflect defects in the apparatus of bipolar budding. Instead, the absence of Ist3 or Bud13 results in a specific defect in the splicing of the MATa 1 pre-mRNA, which encodes a repressor that normally blocks expression of haploid-specific genes in diploid cells. When Mata1 protein is lacking, Axl1, a haploid- specific protein critical for the choice between axial and bipolar budding, is expressed ectopically in diploid cells and disrupts bipolar budding. The involvement of Ist3 and Bud13 in pre-mRNA splicing is by now well known, but the degree of specificity shown here for MATa 1 pre-mRNA, which has no obvious basis in the pre-mRNA structure, is rather surprising in view of current models for the functions of these proteins. Moreover, we found that deletion of PML1, whose product is thought to function together with Ist3 and Bud13 in a three-protein retention-and-splicing (RES) complex, had no detectable effect on the splicing in vivo of either MATa 1 or four other p