Amber mutations in ribosome recycling factors of Escherichia coli and Thermus thermophilus: evidence for C‐terminal modulator element

FEBS Letters

Nakayashiki, Toru¹  Fujiwara, Toshinobu¹  Nakamura, Yoshikazu¹  Ito, Koichi¹ 

[1] Department of Tumor Biology, The Institute of Medical Science, The University of Tokyo, P.O. Takanawa, Tokyo 108-8639, Japan

关键词: Ribosome recycling factor;    Thermus thermophilus ribosome recycling factor;    Amber mutation;    Intergeneric complementation;    Translation termination;    RRF;    ribosome recycling factor;    ecRRF;    Escherichia coli RRF;    ttRRF;    Thermus thermophilus RRF;    RF;    release factor;    RF1;    polypeptide release factor 1;    RF2;    polypeptide release factor 2;    RF3;    polypeptide release factor 3;    Ts;    temperature-sensitive;    Am;    amber;    KmR;    kanamycin resistance;    PCR;    polymerase chain reaction;    IPTG;    isopropyl-1-thio-β-d-galactoside;    PAGE;    polyacrylamide gel electrophoresis;   

DOI  :  10.1016/S0014-5793(99)00302-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Ribosome recycling factor, referred to as RRF, is essential for bacterial growth because of its activity of decomposition of the post-termination complex of the ribosome after release of polypeptides. In this study, we isolated a conditionally lethal amber mutation, named frr-3, in the Escherichia coli RRF gene at amino acid position 161, showing that the truncation of the C-terminal 25 amino acids of RRF is lethal to E. coli. An RRF gene cloned from Thermus thermophilus, whose protein is 44% identical and 68% similar to E. coli RRF, failed to complement the frr-3(Am) allele. However, truncation of the C-terminal five amino acids conferred intergeneric complementation activity on T. thermophilus RRF, demonstrating the modulator activity of the C-terminal tail. Rapid purification of T. thermophilus RRF was achieved by T7-RNA polymerase-driven overexpression for crystallography.

【 授权许可】

Unknown