Intersubunit movement is required for ribosomal translocation

被引:103
作者
Horan, Lucas H.
Noller, Harry F. [1 ]
机构
[1] Univ Calif Santa Cruz, Dept Mol Cell & Dev Biol, Santa Cruz, CA 95064 USA
[2] Univ Calif Santa Cruz, Ctr Mol Biol, RNA, Santa Cruz, CA 95064 USA
关键词
ratchet; tRNA; hybrid state;
D O I
10.1073/pnas.0700762104
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Translocation of tRNA and mRNA during protein synthesis is believed to be coupled to structural changes in the ribosome. The "ratchet model," based on cryo-EM reconstructions of ribosome complexes, invokes relative movement of the 30S and 50S ribosomal subunits in this process; however, evidence that directly demonstrates a requirement for intersubunit movement during translocation is lacking. To address this problem, we created an intersubunit disulfide cross-link to restrict potential movement. The cross-linked ribosomes were unable to carry out polypeptide synthesis; this inhibition was completely reversed upon reduction of the disulfide bridge. In vitro assays showed that the cross-linked ribosomes were specifically blocked in elongation factor G-dependent translocation. These findings show that intersubunit movement is required for ribosomal translocation, accounting for the universal two-subunit architecture of ribosomes.
引用
收藏
页码:4881 / 4885
页数:5
相关论文
共 33 条
[1]   ISOLATION AND FUNCTIONAL-ANALYSIS OF HISTIDINE-TAGGED ELONGATION-FACTOR TU [J].
BOON, K ;
VIJGENBOOM, E ;
MADSEN, LV ;
TALENS, A ;
KRAAL, B ;
BOSCH, L .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1992, 210 (01) :177-183
[2]   TRANSLOCATION IN PROTEIN SYNTHESIS - A HYBRID STRUCTURE MODEL [J].
BRETSCHER, MS .
NATURE, 1968, 218 (5142) :675-+
[3]   TRANSMEMBRANE SIGNALING BY THE ASPARTATE RECEPTOR - ENGINEERED DISULFIDES REVEAL STATIC REGIONS OF THE SUBUNIT INTERFACE [J].
CHERVITZ, SA ;
LIN, CM ;
FALKE, JJ .
BIOCHEMISTRY, 1995, 34 (30) :9722-9733
[4]   A ratchet-like inter-subunit reorganization of the ribosome during translocation [J].
Frank, J ;
Agrawal, RK .
NATURE, 2000, 406 (6793) :318-322
[5]   Catalysis of ribosomal translocation by sparsomycin [J].
Fredrick, K ;
Noller, HF .
SCIENCE, 2003, 300 (5622) :1159-1162
[6]   Accurate translocation of mRNA by the ribosome requires a peptidyl group or its analog on the tRNA moving into the 30S P site [J].
Fredrick, K ;
Noller, HF .
MOLECULAR CELL, 2002, 9 (05) :1125-1131
[7]   STIMULATION OF NON-ENZYMIC R TRANSLOCATION IN RIBOSOMES BY PARA-CHLOROMERCURIBENZOATE [J].
GAVRILOV.LP ;
SPIRIN, AS .
FEBS LETTERS, 1971, 17 (02) :324-&
[8]   High resolution structure of the large ribosomal subunit from a mesophilic Eubacterium [J].
Harms, J ;
Schluenzen, F ;
Zarivach, R ;
Bashan, A ;
Gat, S ;
Agmon, I ;
Bartels, H ;
Franceschi, F ;
Yonath, A .
CELL, 2001, 107 (05) :679-688
[9]   SELECTION OF THE INITIATOR TRANSFER-RNA BY ESCHERICHIA-COLI INITIATION-FACTORS [J].
HARTZ, D ;
MCPHEETERS, DS ;
GOLD, L .
GENES & DEVELOPMENT, 1989, 3 (12A) :1899-1912
[10]   SEPARATION AND AUTORADIOGRAPHY OF MICROGRAM QUANTITIES OF RIBOSOMAL-PROTEINS BY 2-DIMENSIONAL POLYACRYLAMIDE-GEL ELECTROPHORESIS [J].
HOWARD, GA ;
TRAUT, RR .
FEBS LETTERS, 1973, 29 (02) :177-180