PMID:10493856
| Citation |
Brännvall, M and Kirsebom, LA (1999) Manganese ions induce miscleavage in the Escherichia coli RNase P RNA-catalyzed reaction. J. Mol. Biol. 292:53-63 |
|---|---|
| Abstract |
Cleavage by the endoribonuclease RNase P requires the presence of divalent metal ions, of which Mg2+ promotes most efficient cleavage. Here we have studied the importance of there being Mg2+ in RNase P RNA catalysis. It is demonstrated that addition of Mn2+ resulted in a shift of the cleavage site and that this shift was associated with a change in the kinetic constants, in particular kcat. Our data further suggest that the influence of Mn2+ on cleavage site recognition depends on the -1/+73 base-pair in the substrate and the +73/294 base-pair in the RNase P RNA-substrate (RS)-complex. Based on our data we suggest that cleavage in the presence of Mg2+ as the only divalent metal ion proceeds through an intermediate which involves the establishment of the +73/294 base-pair in the RS-complex. By contrast, addition of Mn2+ favours an alternative pathway which results in a shift of the cleavage site. We also studied the influence of Mn2+ on cleavage site recognition and the kinetics of cleavage using various RNase P RNA derivatives carrying substitutions in the region of RNase P RNA that base-pair with the 3' terminal end of the substrate. From these results we conclude that a change in the structure of this RNase P RNA domain influences the involvement of a divalent metal ion(s) in the chemistry of cleavage. |
| Links |
PubMed Online version:10.1006/jmbi.1999.3048 |
| Keywords |
Base Pairing; Base Sequence; Endoribonucleases/metabolism; Escherichia coli/enzymology; Escherichia coli Proteins; Kinetics; Lead/pharmacology; Magnesium/metabolism; Manganese/pharmacology; Molecular Sequence Data; Nucleic Acid Conformation; RNA/metabolism; RNA, Bacterial/metabolism; RNA, Catalytic/metabolism; Ribonuclease P |
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