Base-pairing between 23S rRNA and tRNA in the ribosomal A site

被引:168
作者
Kim, DF [1 ]
Green, R [1 ]
机构
[1] Johns Hopkins Univ, Sch Med, Dept Mol Biol & Genet, Baltimore, MD 21205 USA
关键词
D O I
10.1016/S1097-2765(00)80395-0
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
The aminoacyl (A site) tRNA analog 4-thio-dT-p-C-p-puromycin (s(4)TCPm) photochemically cross-links with high efficiency and specificity to G2553 of 23S rRNA and is peptidyl transferase reactive in its cross-linked state, establishing proximity between the highly conserved 2555 loop in domain V of 23S rRNA and the universally conserved CCA end of tRNA. To test for base-pairing interactions between 23S rRNA and aminoacyl tRNA, site-directed mutations were made at the universally conserved nucleotides U2552 and G2553 of 23S rRNA in both E, coli and B. stearothermophilus ribosomal RNA and incorporated into ribosomes. Mutations at G2553 resulted in dominant growth defects in E. coli and in decreased levels of peptidyl transferase activity in vitro. Genetic analysis in vitro of U2552 and G2553 mutant ribosomes and CCA end mutant tRNA substrates identified a base-pairing interaction between C75 of aminoacyl tRNA and G2553 of 23S rRNA.
引用
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页码:859 / 864
页数:6
相关论文
共 41 条
[1]   Placement of protein and RNA structures into a 5 Å-resolution map of the 50S ribosomal subunit [J].
Ban, N ;
Nissen, P ;
Hansen, J ;
Capel, M ;
Moore, PB ;
Steitz, TA .
NATURE, 1999, 400 (6747) :841-847
[2]   AMINOACYL DERIVATIVES OF NUCLEOSIDES, NUCLEOTIDES AND POLYNUCLEOTIDES .36. THE PEPTIDYLTRANSFERASE CENTER OF ESCHERICHIA-COLI RIBOSOMES - BINDING-SITES FOR THE CYTIDINE 3'-PHOSPHATE RESIDUES OF THE AMINOACYL-TRNA 3'-TERMINUS AND THE INTERRELATIONSHIPS BETWEEN THE ACCEPTOR AND DONOR SITES [J].
BHUTA, P ;
KUMAR, G ;
CHLADEK, S .
BIOCHIMICA ET BIOPHYSICA ACTA, 1982, 696 (02) :208-211
[3]   23S rRNA positions essential for tRNA binding in ribosomal functional sites [J].
Bocchetta, M ;
Xiong, LQ ;
Mankin, AS .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (07) :3525-3530
[4]   X-ray crystal structures of 70S ribosome functional complexes [J].
Cate, JH ;
Yusupov, MM ;
Yusupova, GZ ;
Earnest, TN ;
Noller, HF .
SCIENCE, 1999, 285 (5436) :2095-2104
[5]  
CHLADEK S, 1980, BIOL IMPLICATIONS PR, P149
[6]   SYNTHESIS AND THERMAL MELTING BEHAVIOR OF OLIGOMER-POLYMER COMPLEXES CONTAINING DEFINED LENGTHS OF MISMATCHED DA.DG AND DG.DG NUCLEOTIDES [J].
DODGSON, JB ;
WELLS, RD .
BIOCHEMISTRY, 1977, 16 (11) :2367-2374
[7]   ACTION OF SINGLE-STRAND SPECIFIC NUCLEASES ON MODEL DNA HETERODUPLEXES OF DEFINED SIZE AND SEQUENCE [J].
DODGSON, JB ;
WELLS, RD .
BIOCHEMISTRY, 1977, 16 (11) :2374-2379
[8]  
Fahnestock S, 1974, Methods Enzymol, V30, P554
[9]  
Green J, 1996, AEROSPACE, V23, P2
[10]   Mutations at nucleotides G2251 and U2585 of 23 S rRNA perturb the peptidyl transferase center of the ribosome [J].
Green, R ;
Samaha, RR ;
Noller, HF .
JOURNAL OF MOLECULAR BIOLOGY, 1997, 266 (01) :40-50