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Archaea
Volume 1 (2004), Issue 4, Pages 241-245
http://dx.doi.org/10.1155/2004/903283
Research Article

In vitro selection of an archaeal RNase P RNA mimics natural variation

1Department of Microbiology, North Carolina State University, Raleigh, NC 27695-7615, USA
2V.A. Medical Center, Emory University Medical Research, Rm 5A188, 1670 Clarimont Rd., Decatur, GA 30033, USA

Received 17 September 2003; Accepted 15 October 2003

Copyright © 2004 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Archaeal and bacterial RNase P RNAs are similar in sequence and secondary structure, but in the absence of protein, the archaeal RNAs are much less active and require extreme ionic conditions for activity. To assess how readily the activity of the archaeal RNA alone could be improved by small changes in sequence, in vitro selection was used to generate variants of a Methanobacterium formicicum RNase P RNA: Bacillus subtilus pre-tRNAAsp self-cleaving conjugate RNA. Functional variants were generated with a spectrum of mutations that were predominately consistent with natural variation in this RNA. Variants generated from the selection had cleavage rates comparable to that of wild type; variants with improved cleavage rates or lower ionic requirements were not obtained. This suggests that the RNase P RNAs of Bacteria and Archaea are globally optimized and the basis for the large biochemical differences between these two types of RNase P RNA is distributed in the molecule.