MAPPING THE PATH OF THE NASCENT PEPTIDE-CHAIN THROUGH THE 23S RNA IN THE 50S RIBOSOMAL-SUBUNIT

被引:44
作者
STADE, K [1 ]
JUNKE, N [1 ]
BRIMACOMBE, R [1 ]
机构
[1] MAX PLANCK INST MOLEK GENET AG RIBOSOMEN, D-14195 BERLIN, GERMANY
关键词
D O I
10.1093/nar/23.13.2371
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
Peptides of different lengths encoded by suitable mRNA fragments were biosynthesized in situ on Escherichia coli ribosomes, The peptides carried a diazirine derivative bound to their N-terminal methionine residue, which was photoactivated whilst the peptides were still attached to the ribosome, Subsequently, the sites of photo-cross-linking to 23S RNA were analyzed by our standard procedures, The N-termini of peptides of increasing length became progressively cross-linked to nucleotide 750 (peptides of 6, 9 or 13-15 amino acids), to nucleotide 1614 and concomitantly to a second site between nucleotides 1305 and 1350 (a peptide of 25-26 amino acids), and to nucleotide 91 (a peptide of 29-33 amino acids), Previously we had shown that peptides of 1 or 2 amino acids were cross-linked to nucleotides 2062, 2506 and 2585 within the peptidyl transferase ring, whereas triand tetrapeptides were additionally cross-linked to nucleotides 2609 and 1781, Taken together, the data demonstrate that the path of the nascent peptide chain moves from the peptidyl transferase ring in domain V of the 23S RNA to domain IV, then to domain II, then to domain III, and finally to domain I, These cross-linking results are correlated with other types of topographical data relating to the 50S subunit.
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页码:2371 / 2380
页数:10
相关论文
共 50 条
[1]  
[Anonymous], 1989, MOL CLONING
[2]   4 NEWLY LOCATED PSEUDOURIDYLATE RESIDUES IN ESCHERICHIA-COLI 23S RIBOSOMAL-RNA ARE ALL AT THE PEPTIDYLTRANSFERASE CENTER - ANALYSIS BY THE APPLICATION OF A NEW SEQUENCING TECHNIQUE [J].
BAKIN, A ;
OFENGAND, J .
BIOCHEMISTRY, 1993, 32 (37) :9754-9762
[3]   NASCENT POLYPEPTIDE-CHAINS EMERGE FROM THE EXIT DOMAIN OF THE LARGE RIBOSOMAL-SUBUNIT - IMMUNE MAPPING OF THE NASCENT CHAIN [J].
BERNABEU, C ;
LAKE, JA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1982, 79 (10) :3111-3115
[4]   CONTROLLED PROTEOLYSIS OF NASCENT POLYPEPTIDES IN RAT LIVER CELL FRACTIONS .1. LOCATION OF POLYPEPTIDES WITHIN RIBOSOMES [J].
BLOBEL, G ;
SABATINI, DD .
JOURNAL OF CELL BIOLOGY, 1970, 45 (01) :130-&
[5]   APPLICATION OF 3-[3-(3-(TRIFLUOROMETHYL)DIAZIRIN-3-YL)PHENYL]-2,3-DIHYDROXYPROPIONIC ACID, CARBENE-GENERATING, CLEAVABLE CROSS-LINKING REAGENT FOR PHOTOAFFINITY-LABELING [J].
BOCHKARIOV, DE ;
KOGON, AA .
ANALYTICAL BIOCHEMISTRY, 1992, 204 (01) :90-95
[6]   PRIMARY AND SECONDARY STRUCTURES OF ESCHERICHIA-COLI MRE-600-23S RIBOSOMAL-RNA - COMPARISON WITH MODELS OF SECONDARY STRUCTURE FOR MAIZE CHLOROPLAST 23S RIBOSOMAL-RNA AND FOR LARGE PORTIONS OF MOUSE AND HUMAN 16S MITOCHONDRIAL RIBOSOMAL-RNAS [J].
BRANLANT, C ;
KROL, A ;
MACHATT, MA ;
POUYET, J ;
EBEL, JP ;
EDWARDS, K ;
KOSSEL, H .
NUCLEIC ACIDS RESEARCH, 1981, 9 (17) :4303-4324
[7]   CLUSTERING OF MODIFIED NUCLEOTIDES AT THE FUNCTIONAL CENTER OF BACTERIAL RIBOSOMAL-RNA [J].
BRIMACOMBE, R ;
MITCHELL, P ;
OSSWALD, M ;
STADE, K ;
BOCHKARIOV, D .
FASEB JOURNAL, 1993, 7 (01) :161-167
[8]  
BRIMACOMBE R, 1990, RIBOSOME, P93
[9]  
BRIMACOMBE R, 1995, EUR J BIOCHEM, V230, P365
[10]  
CANTOR CR, 1974, RIBOSOMES, P573