O'Connor J P, Peebles C L
Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260.
Mol Cell Biol. 1992 Sep;12(9):3843-56. doi: 10.1128/mcb.12.9.3843-3856.1992.
We have identified an essential Saccharomyces cerevisiae gene, PTA1, that affects pre-tRNA processing. PTA1 was initially defined by a UV-induced mutation, pta1-1, that causes the accumulation of all 10 end-trimmed, intron-containing pre-tRNAs and temperature-sensitive but osmotic-remedial growth. pta1-1 does not appear to be an allele of any other known gene affecting pre-tRNA processing. Extracts prepared from pta1-1 strains had normal pre-tRNA splicing endonuclease activity. pta1-1 was suppressed by the ochre suppressor tRNA gene SUP11, indicating that the pta1-1 mutation creates a termination codon within a protein reading frame. The PTA1 gene was isolated from a genomic library by complementation of the pta1-1 growth defect. Episome-borne PTA1 directs recombination to the pta1-1 locus. PTA1 has been mapped to the left arm of chromosome I near CDC24; the gene was sequenced and could encode a protein of 785 amino acids with a molecular weight of 88,417. No other protein sequences similar to that of the predicted PTA1 gene product have been identified within the EMBL or GenBank data base. Disruption of PTA1 near the carboxy terminus of the putative open reading frame was lethal. Possible functions of the PTA1 gene product are discussed.
我们鉴定出了酿酒酵母中的一个必需基因PTA1,它影响前体tRNA的加工过程。PTA1最初是由紫外线诱导的突变pta1-1所定义的,该突变导致所有10种经过末端修剪、含有内含子的前体tRNA积累,并导致温度敏感但渗透压可补救的生长。pta1-1似乎不是任何其他已知影响前体tRNA加工基因的等位基因。从pta1-1菌株制备的提取物具有正常的前体tRNA剪接内切核酸酶活性。pta1-1被赭石抑制tRNA基因SUP11抑制,这表明pta1-1突变在蛋白质阅读框内产生了一个终止密码子。通过互补pta1-1的生长缺陷,从基因组文库中分离出了PTA1基因。附加型携带的PTA1引导重组至pta1-1位点。PTA1已被定位到靠近CDC24的第一条染色体的左臂;该基因已被测序,可能编码一个由785个氨基酸组成、分子量为88417的蛋白质。在EMBL或GenBank数据库中未发现与预测的PTA1基因产物相似的其他蛋白质序列。在假定开放阅读框的羧基末端附近破坏PTA1是致死的。文中还讨论了PTA1基因产物可能的功能。