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
收藏
页码:859 / 864
页数:6
相关论文
共 41 条
[11]   Ribosome-catalyzed peptide-bond formation with an a-site substrate covalently linked to 23S ribosomal RNA [J].
Green, R ;
Switzer, C ;
Noller, HF .
SCIENCE, 1998, 280 (5361) :286-289
[12]   Reconstitution of functional 50S ribosomes from in vitro transcripts of Bacillus stearothermophilus 23S rRNA [J].
Green, R ;
Noller, HF .
BIOCHEMISTRY, 1999, 38 (06) :1772-1779
[13]   Ribosomes and translation [J].
Green, R ;
Noller, HF .
ANNUAL REVIEW OF BIOCHEMISTRY, 1997, 66 :679-716
[14]   A COMPILATION OF LARGE SUBUNIT (23S-LIKE AND 23S-LIKE) RIBOSOMAL-RNA STRUCTURES [J].
GUTELL, RR ;
SCHNARE, MN ;
GRAY, MW .
NUCLEIC ACIDS RESEARCH, 1992, 20 :2095-2109
[15]   Surprising fidelity of template-directed chemical ligation of oligonucleotides [J].
James, KD ;
Ellington, AD .
CHEMISTRY & BIOLOGY, 1997, 4 (08) :595-605
[16]   Movement of the 3'-end of tRNA through the peptidyl transferase centre and its inhibition by antibiotics [J].
Kirillov, S ;
Porse, BT ;
Vester, B ;
Woolley, P ;
Garrett, RA .
FEBS LETTERS, 1997, 406 (03) :223-233
[17]  
LIEBERMAN KR, 1994, J BIOL CHEM, V269, P16163
[18]  
LIEBERMAN KR, 1995, PROG NUCLEIC ACID RE, V50, P1, DOI 10.1016/S0079-6603(08)60809-0
[19]   Nucleotides in 16S rRNA protected by the association of 30 S and 50 S ribosomal subunits [J].
Merryman, C ;
Moazed, D ;
McWhirter, J ;
Noller, HF .
JOURNAL OF MOLECULAR BIOLOGY, 1999, 285 (01) :97-105
[20]   Nucleotides in 23 S rRNA protected by the association of 30 S and 50 S ribosomal subunits [J].
Merryman, C ;
Moazed, D ;
Daubresse, G ;
Noller, HF .
JOURNAL OF MOLECULAR BIOLOGY, 1999, 285 (01) :107-113