Examining the bases of the J3/4 domain of Escherichia coli ribonuclease P.
We prepared several mutants of the J3/4 and P4 domains of Escherichia coli ribonuclease P (RNase P): A62G, A62U, G63C/G64C, A65G, A67G, U69A, U69G, U69C, U69Delta, and U69UU. Comparison of the ribozyme and holo enzyme reactions at various concentrations of magnesium ions showed that the presence of a bulge at U69 in the P4 domain was important in the holo enzyme. The results also showed that the conserved bases G63 and G64 in the J3/4 domain were important for efficient ribozyme reactions but were replaceable in the presence of the protein component. Our data showed that the bases in the J3/4 and P4 domains displayed different responses to the metal ions that were affected by the presence of the protein component.
Tanaka
Terumichi
T
Division of Bioscience and Biotechnology, Department of Ecological Engineering, Toyohashi University of Technology, Toyohashi, Japan. tanakat@eco.tut.ac.jp
Ando
Tomoaki
T
Haga
Shinnosuke
S
Kikuchi
Yo
Y
D001483:Base Sequence; D029968:Escherichia coli Proteins / Q000737:chemistry; D008274:Magnesium; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D009690:Nucleic Acid Conformation; D017434:Protein Structure, Tertiary; D016337:RNA, Catalytic / Q000378:metabolism; D043262:Ribonuclease P / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D029968:Escherichia coli Proteins; D016337:RNA, Catalytic; D043262:Ribonuclease P; D008274:Magnesium