Colicin E5 ribonuclease domain cleaves Saccharomyces cerevisiae tRNAs leading to impairment of the cell growth.
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.
Ogawa
Tetsuhiro
T
Department of Biotechnology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Hidaka
Makoto
M
Kohno
Kenji
K
Masaki
Haruhiko
H
D001483:Base Sequence; D003087:Colicins / Q000737:chemistry* / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D050296:Microbial Viability; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D017434:Protein Structure, Tertiary; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012260:Ribonucleases / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000378:metabolism*; D013379:Substrate Specificity; D014170:Transformation, Genetic
D003087:Colicins; D011993:Recombinant Fusion Proteins; C096880:colicin 5 protein, E coli; D012343:RNA, Transfer; D012260:Ribonucleases