Maekawa H, Nakagawa T, Uno Y, Kitamura K, Shimoda C
Department of Biology, Faculty of Science, Osaka City University, Japan.
Mol Gen Genet. 1994 Sep 1;244(5):456-64. doi: 10.1007/BF00583896.
When the fission yeast Schizosaccharomyces pombe is starved for nitrogen, the cells are arrested in the G1 phase, enter the G0 phase and initiate sexual development. The ste13 mutant, however, fails to undergo a G1 arrest when starved for nitrogen and since this mutant phenotype is not suppressed by a mutation in adenylyl cyclase (cyr1), it would appear that ste13+ either acts independently of the decrease in the cellular cAMP level induced by starvation for nitrogen, or functions downstream of this controlling event. We have used functional complementation to clone the ste13+ gene from an S. pombe genomic library and show that its disruption is not lethal, indicating that, while the gene is required for sexual development, it is not essential for cell growth. Nucleotide sequencing predicts that ste13+ should encode a protein of 485 amino acids in which the consensus motifs of ATP-dependent RNA helicases of the DEAD box family are completely conserved. Point mutations introduced into these consensus motifs abolished the ste13+ functions. The predicted Ste13 protein is 72% identical to the Drosophila melanogaster Me31B protein over a stretch of 391 amino acids. ME31B is a developmentally regulated gene that is expressed preferentially in the female germline and may be required for oogenesis. Expression of ME31B cDNA in S. pombe suppresses the ste13 mutation. These two evolutionarily conserved genes encoding putative RNA helicases may play a pivotal role in sexual development.
当裂殖酵母粟酒裂殖酵母缺乏氮源时,细胞会停滞在G1期,进入G0期并启动有性发育。然而,ste13突变体在缺乏氮源时无法发生G1期停滞,并且由于该突变体表型不会被腺苷酸环化酶(cyr1)的突变所抑制,因此似乎ste13+要么独立于因缺乏氮源而导致的细胞内cAMP水平降低发挥作用,要么在这一调控事件的下游起作用。我们利用功能互补从粟酒裂殖酵母基因组文库中克隆了ste13+基因,并表明其缺失并非致命,这表明虽然该基因是有性发育所必需的,但对细胞生长并非必不可少。核苷酸测序预测ste13+应编码一个485个氨基酸的蛋白质,其中DEAD盒家族的ATP依赖性RNA解旋酶的共有基序完全保守。引入这些共有基序的点突变消除了ste13+的功能。预测的Ste13蛋白在一段391个氨基酸的区域内与黑腹果蝇的Me31B蛋白有72%的同一性。ME31B是一个受发育调控的基因,在雌性生殖系中优先表达,可能是卵子发生所必需的。在粟酒裂殖酵母中表达ME31B cDNA可抑制ste13突变。这两个编码推定RNA解旋酶的进化保守基因可能在有性发育中起关键作用。