TY - JOUR
T1 - Analysis of the functional role of conserved residues in the protein subunit of ribonuclease P from Escherichia coli
AU - Gopalan, Venkat
AU - Baxevanis, Andreas D.
AU - Landsman, David
AU - Altman, Sidney
N1 - Funding Information:
We are grateful to members of our laboratory, especially Drs Cecilia Guerrier-Takada, Paul Eder and Urban Lundberg, for helpful discussions. Research in the laboratory of S.A. is supported by National Institutes of Health grant GM19422. V.G. was funded by postdoctoral fellowships from the Patrick and Catherine Donaghue Foundation for Medical Research and the Anna Fuller Fund for Cancer Research.
PY - 1997/4/11
Y1 - 1997/4/11
N2 - The processing of precursor tRNAs and some other small cellular RNAs by M1 RNA, the catalytic subunit of Escherichia coli ribonuclease P, is accelerated by C5 protein (the protein cofactor) both in vitro and in vivo. In an effort to understand the mechanism by which the protein cofactor promotes and stabilizes certain conformations of M1 RNA that are most efficient for RNase P catalysis, we have used site-directed mutagenesis to generate mutant derivatives of C5 protein and assessed their ability to promote RNase P catalysis in vivo and in vitro. Our results indicate that certain conserved hydrophobic and basic residues in C5 protein are important for its function and that single amino acid residue changes in C5 protein can alter the substrate specificity of the RNase P holoenzyme.
AB - The processing of precursor tRNAs and some other small cellular RNAs by M1 RNA, the catalytic subunit of Escherichia coli ribonuclease P, is accelerated by C5 protein (the protein cofactor) both in vitro and in vivo. In an effort to understand the mechanism by which the protein cofactor promotes and stabilizes certain conformations of M1 RNA that are most efficient for RNase P catalysis, we have used site-directed mutagenesis to generate mutant derivatives of C5 protein and assessed their ability to promote RNase P catalysis in vivo and in vitro. Our results indicate that certain conserved hydrophobic and basic residues in C5 protein are important for its function and that single amino acid residue changes in C5 protein can alter the substrate specificity of the RNase P holoenzyme.
KW - RNase P protein subunit
KW - Site-directed mutagenesis
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U2 - 10.1006/jmbi.1997.0906
DO - 10.1006/jmbi.1997.0906
M3 - Article
C2 - 9135114
AN - SCOPUS:0031564661
SN - 0022-2836
VL - 267
SP - 818
EP - 829
JO - Journal of molecular biology
JF - Journal of molecular biology
IS - 4
ER -