Cleavage mediated by the catalytic domain of bacterial RNase P RNA
2012 (English)In: Journal of Molecular Biology, ISSN 0022-2836, E-ISSN 1089-8638, Vol. 422, no 2, 204-214 p.Article in journal (Refereed) Published
As for other RNA molecules RNase P RNA (RPR) is composed of domains and these have different functions. Here we provide data demonstrating that the catalytic (C) domain of Escherichia coli (Eco) RPR when separated from the specificity (S) domain mediates cleavage using various model hairpin loop substrates. Compared to full-size Eco RPR the rate of cleavage for the truncated RPR (CP RPR) was reduced 30- to 13000-fold. We provide data that the magnitude of reduction in rate is substrate dependent and that the structural architecture of the -1/+73 plays a significant role where a C-1/G+73 pair had the most dramatic effect on the rate. Substitution of A248 (E. coli numbering), which is positioned near the cleavage site in the RNase P-substrate complex, with G in the CP RPR resulted in 30-fold rate improvement while strengthening the interaction between the RPR and the 3' end of the substrate only had a modest effect. Interestingly, while deleting the S-domain gave a reduction in the rate it resulted in a less erroneous RPR with respect to cleavage site selection. These data will be dicussed in view of our current understanding of the coupling between the distal interaction between the S-domain and events at the active site and in an evolutionary perspective.
Place, publisher, year, edition, pages
2012. Vol. 422, no 2, 204-214 p.
RNase P/Ribozyme/Divalent metal ions/tRNA precursors/tRNA processing
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:uu:diva-159311DOI: 10.1016/j.jmb.2012.05.020ISI: 000308681000005OAI: oai:DiVA.org:uu-159311DiVA: diva2:444087