Stimulation of the GTPase activity of translation elongation factor G by ribosomal protein L7/12

被引:76
作者
Savelsbergh, A [1 ]
Mohr, D [1 ]
Wilden, B [1 ]
Wintermeyer, W [1 ]
Rodnina, MV [1 ]
机构
[1] Univ Witten Herdecke, Inst Mol Biol, D-58448 Witten, Germany
关键词
D O I
10.1074/jbc.275.2.890
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
Elongation factors (EFs) Tu and G are GTPases that have important functions in protein synthesis. The low intrinsic GTPase activity of both factors is strongly stimulated on the ribosome by unknown mechanisms. Here we report that isolated ribosomal protein L7/12 strongly stimulates GTP hydrolysis by EF-G, but not by EF-Tu, indicating a major contribution of L7/12 to GTPase activation of EF-G on the ribosome. The effect is due to the acceleration of the catalytic step because the rate of GDP-GTP exchange on EF-G, as measured by rapid kinetics, is much faster than the steady-state GTPase rate. The unique, highly conserved arginine residue in the C-terminal domain of L7/12 is not essential for the activation, excluding an "arginine finger"-type mechanism. L7/12 appears to function by stabilizing the GTPase transition state of EF-G.
引用
收藏
页码:890 / 894
页数:5
相关论文
共 49 条
[1]   Visualization of elongation factor G on the Escherichia coli 70S ribosome:: The mechanism of translocation [J].
Agrawal, RK ;
Penczek, P ;
Grassucci, RA ;
Frank, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (11) :6134-6138
[2]   EF-G-dependent GTP hydrolysis induces translocation accompanied by large conformational changes in the 70S ribosome [J].
Agrawal, RK ;
Heagle, AB ;
Penczek, P ;
Grassucci, RA ;
Frank, J .
NATURE STRUCTURAL BIOLOGY, 1999, 6 (07) :643-647
[3]   Confirmation of the arginine-finger hypothesis for the GAP-stimulated GTP-hydrolysis reaction of Ras [J].
Ahmadian, MR ;
Stege, P ;
Scheffzek, K ;
Wittinghofer, A .
NATURE STRUCTURAL BIOLOGY, 1997, 4 (09) :686-689
[4]   Conformational independence of N- and C-domains in ribosomal protein L7/L12 and in the complex with protein L10 [J].
Bocharov, EV ;
Gudkov, AT ;
Budovskaya, EV ;
Arseniev, AS .
FEBS LETTERS, 1998, 423 (03) :347-350
[5]  
BOURNE HR, 1991, NATURE, V349, P117, DOI 10.1038/349117a0
[6]   STRUCTURES OF ACTIVE CONFORMATIONS OF G(I-ALPHA-1) AND THE MECHANISM OF GTP HYDROLYSIS [J].
COLEMAN, DE ;
BERGHUIS, AM ;
LEE, E ;
LINDER, ME ;
GILMAN, AG ;
SPRANG, SR .
SCIENCE, 1994, 265 (5177) :1405-1412
[7]   Crystal structure of a conserved ribosomal protein-RNA complex [J].
Conn, GL ;
Draper, DE ;
Lattman, EE ;
Gittis, AG .
SCIENCE, 1999, 284 (5417) :1171-1174
[8]   GUANOSINETRIPHOSPHATASE ACTIVITY DEPENDENT ON ELONGATION FACTOR-TU AND RIBOSOMAL-PROTEIN L7-L12 [J].
DONNER, D ;
VILLEMS, R ;
LILJAS, A ;
KURLAND, CG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1978, 75 (07) :3192-3195
[9]  
GALE EF, 1981, ANTIBIOTIC INHIBITOR, P402
[10]   Structural and functional analysis of the ARF1-ARFGAP complex reveals a role for coatomer in GTP hydrolysis [J].
Goldberg, J .
CELL, 1999, 96 (06) :893-902