Ogawa Tetsuhiro, Hidaka Makoto, Kohno Kenji, Masaki Haruhiko
Department of Biotechnology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
J Biochem. 2009 Apr;145(4):461-6. doi: 10.1093/jb/mvp004. Epub 2009 Jan 16.
Colicin E5 is a ribonuclease that specifically cleaves tRNA(Tyr), tRNA(His), tRNA(Asn) and tRNA(Asp) of sensitive Escherichia coli cells by recognizing their anticodon sequences. Since all organisms possess universal anticodons of these tRNAs, colicin E5 was expected to potentially cleave eukaryotic tRNAs. Here, we expressed the active domain of colicin E5 (E5-CRD) in Saccharomyces cerevisiae and investigated its effects on growth. E5-CRD impaired growth of host cells by cleaving tRNA(Tyr), tRNA(His), tRNA(Asn) and tRNA(Asp) in S. cerevisiae, which is the same repertoire as that in E. coli. This activity of E5-CRD was inhibited by the co-expression of its cognate inhibitor (ImmE5). Notably, the growth impairment by E5-CRD was reversible; cells restored the colony-forming activity after suppression of the E5-CRD expression. This seems different from the sharp killing effect of E5-CRD on E. coli. These results may provide insights into the role and behaviour of cytosolic tRNAs on cell growth and proliferation.
大肠杆菌素E5是一种核糖核酸酶,它通过识别敏感大肠杆菌细胞的反密码子序列,特异性地切割这些细胞中的tRNA(Tyr)、tRNA(His)、tRNA(Asn)和tRNA(Asp)。由于所有生物体都拥有这些tRNA的通用反密码子,因此预计大肠杆菌素E5可能会切割真核生物的tRNA。在这里,我们在酿酒酵母中表达了大肠杆菌素E5的活性结构域(E5-CRD),并研究了其对生长的影响。E5-CRD通过切割酿酒酵母中的tRNA(Tyr)、tRNA(His)、tRNA(Asn)和tRNA(Asp)来损害宿主细胞的生长,这与在大肠杆菌中的情况相同。E5-CRD的这种活性受到其同源抑制剂(ImmE5)共表达的抑制。值得注意的是,E5-CRD对生长的损害是可逆的;在抑制E5-CRD表达后,细胞恢复了集落形成活性。这似乎与E5-CRD对大肠杆菌的急剧杀伤作用不同。这些结果可能为胞质tRNA在细胞生长和增殖中的作用及行为提供见解。