Kishida M, Nagai T, Nakaseko Y, Shimoda C
Department of Biology, Faculty of Science, Osaka City University, Japan.
Curr Genet. 1994 Jun;25(6):497-503. doi: 10.1007/BF00351668.
The mes1+ gene of the fission yeast Schizosaccharomyces pombe is essential for the second meiotic division. We have cloned a 1.1-kb HindIII fragment containing mes1+ by complementation from an S. pombe genomic library. Sequencing of the genomic and cDNA fragments indicates the existence of one small intron of 75 nucleotides, although both the 5'(G/GTTAGT) and 3'(CAG/T) intron-exon junctions deviate from the consensus sequences proposed for S. pombe. The putative translation product of the mature mes1+ mRNA is a 11-kDa protein of 101 amino acids which has no significant homology to any previously-reported proteins. Disruption of mes1 has no effect on cell growth but causes an arrest of meiosis before the second meiotic division. Northern-blot analysis revealed that mes1+ was preferentially transcribed under conditions of nitrogen starvation. When a h90 homothallic strain was shifted to a nitrogen-deficient medium, a pre-mRNA accumulated and then was gradually processed to generate a mature mRNA. This splicing did not occur in either a heterothallic haploid strain or in a homothallic mei2 mutant strain which was defective in the initiation of meiosis. Expression of the first exon alone was not able to suppress the mes1 null allele. These results indicate that mes1+ is required for the completion of meiosis, that splicing is required for the function of the mes1+ gene, and that this splicing requires the function of the mei2+ product.
裂殖酵母粟酒裂殖酵母的mes1+基因对第二次减数分裂至关重要。我们通过从粟酒裂殖酵母基因组文库中互补克隆了一个包含mes1+的1.1 kb HindIII片段。基因组和cDNA片段的测序表明存在一个75个核苷酸的小内含子,尽管5'(G/GTTAGT)和3'(CAG/T)内含子-外显子连接均偏离了为粟酒裂殖酵母提出的共有序列。成熟mes1+ mRNA的推定翻译产物是一种由101个氨基酸组成的11 kDa蛋白质,与任何先前报道的蛋白质均无明显同源性。mes1的破坏对细胞生长没有影响,但会导致减数分裂在第二次减数分裂之前停滞。Northern印迹分析显示,mes1+在氮饥饿条件下优先转录。当h90同宗配合菌株转移到缺氮培养基中时,一种前体mRNA积累,然后逐渐加工产生成熟mRNA。这种剪接在异宗配合单倍体菌株或在减数分裂起始有缺陷的同宗配合mei2突变菌株中均未发生。仅第一个外显子的表达不能抑制mes1无效等位基因。这些结果表明,mes1+是减数分裂完成所必需的,剪接是mes1+基因功能所必需的,并且这种剪接需要mei2+产物的功能。