The detection of natural opal suppressor seryl‐tRNA in Escherichia coli by the dot blot hybridization and its phosphorylation by a tRNA kinase

FEBS Letters

Hitaka, Teruaki²  Sukenaga, Yoshikazu¹  Mizutani, Takaharu²  Maruyama, Naosuke² 

[1] Research Laboratory, Nippon Kayaku Co. Ltd, 3-31 Shimo, Kita-ku, Tokyo 115, Japan;Faculty of Pharmaceutical Sciences, Nagoya City University, Mizuho-ku, Nagoya 467 Japan

关键词: Suppressor tRNA;    Formate dehydrogenase;    Selenocysteine;    Opal termination codon;    tRNA;    phosphoseryl-;   

DOI  :  10.1016/0014-5793(89)81367-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

It was believed that there was no natural suppressor tRNA in Escherichia coli, however, it has been suggested that selC, relating to the synthesis of formate dehydrogenase of a selenoprotein [(1988) Nature 331, 723–725], codes for tRNA, even though the presence of tRNA has not yet been demonstrated. We detected the product of selC in the tRNA preparation of the E. coli MC 4100 strain by the dot blot hybridization method with a DNA probe (ACCGCTGGCGGC) corresponding to the extra arm of selC tRNA. Two hybridization peaks were found in the chromatographic pattern from Sephadex A50. The amount of tRNA was estimated to be about 0.03% of the total tRNA. The suppressor [³H]seryl-tRNA was phosphorylated by a tRNA kinase in E. coli B. These results suggest that the opal suppressor seryl-tRNA in E. coli should be converted to selenocysteyl-tRNA through phosphoseryl-tRNA, and occurs in vertebrates as a general phenomenon.

【 授权许可】

Unknown