Localization of L11 protein on the ribosome and elucidation of its involvement in EF-G-dependent translocation

被引:94
作者
Agrawal, RK
Linde, J
Sengupta, J
Nierhaus, KH
Frank, J
机构
[1] New York State Dept Hlth, Wadsworth Ctr Labs & Res, Hlth Res Inc, Howard Hughes Med Inst, Albany, NY 12201 USA
[2] SUNY Albany, Dept Biomed Sci, Albany, NY 12201 USA
[3] Max Planck Inst Mol Genet, D-14195 Berlin, Germany
关键词
elongation factor release; thiostrepton; translation;
D O I
10.1006/jmbi.2001.4907
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
L11 protein is located at the base of the L7/LI2 stalk of the 50 S subunit of the Escherichia coli ribosome. Because of the flexible nature of the region, recent X-ray crystallographic studies of the 50 S subunit failed to locate the N-terminal domain of the protein. We have determined the position of the complete L11 protein by comparing a three-dimensional cryo-EM reconstruction of the 70 S ribosome, isolated from a mutant lacking ribosomal protein L11, with the three-dimensional map of the wild-type ribosome. Fitting of the X-ray coordinates of L11-23 S RNA complex and EF-G into the cryo-EM maps combined with molecular modeling, reveals that, following EF-G-dependent GTP hydrolysis, domain V of EF-G intrudes into the cleft between the 23 S ribosomal RNA and the N-terminal domain of L11 (where the antibiotic thiostrepton binds), causing the N-terminal domain to move and thereby inducing the formation of the arc-like connection with the G' domain of EF-G. The results provide a new insight into the mechanism of EF-G-depenclent translocation.
引用
收藏
页码:777 / 787
页数:11
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