Mutational analysis of the length of the J3/4 domain of Escherichia coli ribonuclease P ribozyme

We prepared a series of length variants of the J3/4 domain of Escherichia coli ribonuclease P (RNase P) ribozyme: the four-base long J3/4 domain (A62G63G64A65) was replaced with GGA (denoted DeltaA), GA (DeltaAG), A (DeltaAGG), AAGGA (sumA), AAAGGA (sumAA), and AAAAGGA (sumAAA). The results indicate...

Full description

Saved in:
Bibliographic Details
Published in:Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2004-12, Vol.68 (12), p.2630-2632
Main Authors: Haga, S. (Toyohashi Univ. of Technology, Aichi (Japan)), Tanaka, T, Kikuchi, Y
Format: Article
Language:English
Subjects:
Biological and medical sciences
BIOSYNTHESIS
Catalysis
Conserved Sequence
DEGRADATION
ENZYME ACTIVITY
ESCHERICHIA COLI
Escherichia coli - genetics
Fundamental and applied biological sciences. Psychology
J3/4 domain
Kinetics
M1 RNA
MAGNESIUM
Magnesium - pharmacology
Mutagenesis, Site-Directed
NUCLEOTIDE SEQUENCE
Ribonuclease P
ribonuclease P (RNase P)
RIBONUCLEASES
ribozyme
RNA, Catalytic - genetics
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We prepared a series of length variants of the J3/4 domain of Escherichia coli ribonuclease P (RNase P) ribozyme: the four-base long J3/4 domain (A62G63G64A65) was replaced with GGA (denoted DeltaA), GA (DeltaAG), A (DeltaAGG), AAGGA (sumA), AAAGGA (sumAA), and AAAAGGA (sumAAA). The results indicated that truncating and inserting operations of the J3/4 domain drastically reduced ribozyme activity (WT>>sumAA>sumA>sumAAA>>DeltaAG>DeltaA, DeltaAGG), but did not affect the cleavage site selection of a substrate by the ribozyme. The reduced ribozyme activity of each mutant was rescued to some extent by the addition of a high concentration of magnesium ions. Our data indicate that the conserved AGGA sequence was important for efficient ribozyme reactions, and suggested that the length mutations affected ribozyme activity through metal ion binding steps.
ISSN:0916-8451
1347-6947
DOI:10.1271/bbb.68.2630