PRODUCT RELEASE IS A RATE-LIMITING STEP DURING CLEAVAGE BY THE CATALYTIC RNA SUBUNIT OF ESCHERICHIA-COLI RNASE-P

被引:85
作者
TALLSJO, A [1 ]
KIRSEBOM, LA [1 ]
机构
[1] BIOMED CTR,DEPT MICROBIOL,BOX 581,S-75123 UPPSALA,SWEDEN
关键词
D O I
10.1093/nar/21.1.51
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 kinetic constants for cleavage of the tRNA(Tyr)Su3 precursor by the M1 RNA of E.coli RNase P were determined in the absence and presence of the C5 protein under single and multiple (steady state) turnover conditions. The rate constant of cleavage in the reaction catalyzed by M1 RNA alone was 5 times higher in single turnover than in multiple turnovers, suggesting that a rate-limiting step is product release. Cleavage by M1 RNA alone and by the holoenzyme under identical buffer conditions demonstrated that C5 facilitated product release. Addition of different product-like molecules under single turnover reaction conditions inhibited cleavage both in the absence and presence of C5. In the presence of C5, the K(i) value for matured tRNA was approximately 20 times higher than in its absence, suggesting that C5 also reduces the interaction between the 5'-matured tRNA and the enzyme. In a growing cell the number of tRNA molecules is approximately 1000 times higher than the number of RNase P molecules. A 100-fold excess of matured tRNA over enzyme clearly inhibited cleavage in vitro. We discuss the possibility that RNase P is involved in the regulation of tRNA expression under certain growth conditions.
引用
收藏
页码:51 / 57
页数:7
相关论文
共 19 条
[1]   M1-RNA, THE RNA SUBUNIT OF ESCHERICHIA-COLI RIBONUCLEASE-P, CAN UNDERGO A PH-SENSITIVE CONFORMATIONAL CHANGE [J].
ALTMAN, S ;
GUERRIERTAKADA, C .
BIOCHEMISTRY, 1986, 25 (06) :1205-1208
[2]   ENZYMATIC CLEAVAGE OF RNA BY RNA [J].
ALTMAN, S ;
BAER, M ;
GUERRIERTAKADA, C ;
VIOQUE, A .
TRENDS IN BIOCHEMICAL SCIENCES, 1986, 11 (12) :515-518
[3]   CHARACTERIZATION INVITRO OF THE DEFECT IN A TEMPERATURE-SENSITIVE MUTANT OF THE PROTEIN SUBUNIT OF RNASE-P FROM ESCHERICHIA-COLI [J].
BAER, MF ;
WESOLOWSKI, D ;
ALTMAN, S .
JOURNAL OF BACTERIOLOGY, 1989, 171 (12) :6862-6866
[4]  
DIJK J, 1974, J BIOL CHEM, V249, P645
[5]   NOVEL REACTIONS OF RNAASE-P WITH A TRANSFER-RNA-LIKE STRUCTURE IN TURNIP YELLOW MOSAIC-VIRUS RNA [J].
GUERRIERTAKADA, C ;
VANBELKUM, A ;
PLEIJ, CWA ;
ALTMAN, S .
CELL, 1988, 53 (02) :267-272
[6]   THE RNA MOIETY OF RIBONUCLEASE-P IS THE CATALYTIC SUBUNIT OF THE ENZYME [J].
GUERRIERTAKADA, C ;
GARDINER, K ;
MARSH, T ;
PACE, N ;
ALTMAN, S .
CELL, 1983, 35 (03) :849-857
[7]   CATALYSIS OF RNA CLEAVAGE BY THE TETRAHYMENA-THERMOPHILA RIBOZYME .2. KINETIC DESCRIPTION OF THE REACTION OF AN RNA SUBSTRATE THAT FORMS A MISMATCH AT THE ACTIVE-SITE [J].
HERSCHLAG, D ;
CECH, TR .
BIOCHEMISTRY, 1990, 29 (44) :10172-10180
[8]   REACTION INVITRO OF SOME MUTANTS OF RNASE-P WITH WILD-TYPE AND TEMPERATURE-SENSITIVE SUBSTRATES [J].
KIRSEBOM, LA ;
ALTMAN, S .
JOURNAL OF MOLECULAR BIOLOGY, 1989, 207 (04) :837-840
[9]   THE KINETICS AND SPECIFICITY OF CLEAVAGE BY RNASE-P IS MAINLY DEPENDENT ON THE STRUCTURE OF THE AMINO-ACID ACCEPTOR STEM [J].
KIRSEBOM, LA ;
SVARD, SG .
NUCLEIC ACIDS RESEARCH, 1992, 20 (03) :425-432
[10]   OLIGORIBONUCLEOTIDE SYNTHESIS USING T7 RNA-POLYMERASE AND SYNTHETIC DNA TEMPLATES [J].
MILLIGAN, JF ;
GROEBE, DR ;
WITHERELL, GW ;
UHLENBECK, OC .
NUCLEIC ACIDS RESEARCH, 1987, 15 (21) :8783-8798