Kumar Chitranshu, Sharma Rakesh, Bachhawat Anand Kumar
Institute of Microbial Technology, Chandigarh 160 036, India.
Yeast. 2003 Jul 30;20(10):857-63. doi: 10.1002/yea.1012.
The ECM38 gene encodes the gamma-glutamyl transpeptidase enzyme, an enzyme involved in glutathione turnover. The enzyme was found to be present in the S288C strain, BY4742, but absent in another widely used strain congenic to S288C, YPH499. Cloning and sequencing the genes from these yeasts indicated the presence of 11 single nucleotide polymorphisms in the coding region and eight single nucleotide polymorphisms in the promoter region of the ECM38 gene of YPH499 (but none in that of BY4742). One of the SNPs in the ECM38 ORF led to a G --> D conversion in a region conserved in all gamma-GT enzymes and was found to be responsible for the loss of activity in this strain. The presence of gamma-GT activity in other YPH strains led us to trace the origins of the polymorphisms in YPH499. Our results indicated that among the progenitor strains, YPH1 and YPH2, YPH1 carried the polymorphisms seen in YPH499 and also lacked the gamma-GT activity. The implications of these results for the use of these widely used S288C strains and the origin of these single nucleotide polymorphisms are presented.
ECM38基因编码γ-谷氨酰转肽酶,该酶参与谷胱甘肽的代谢周转。研究发现,在S288C菌株BY4742中存在这种酶,但在另一种与S288C同基因的广泛使用的菌株YPH499中却不存在。对这些酵母的基因进行克隆和测序表明,YPH499的ECM38基因编码区存在11个单核苷酸多态性,启动子区存在8个单核苷酸多态性(而BY4742的该基因不存在单核苷酸多态性)。ECM38开放阅读框中的一个单核苷酸多态性导致了所有γ-GT酶保守区域的G→D转换,并且发现该转换是导致该菌株活性丧失的原因。其他YPH菌株中γ-GT活性的存在促使我们追溯YPH499中多态性的起源。我们的结果表明,在祖代菌株YPH1和YPH2中,YPH1携带了YPH499中出现的多态性,并且也缺乏γ-GT活性。本文介绍了这些结果对于这些广泛使用的S288C菌株的使用以及这些单核苷酸多态性起源的意义。