title: In vitro selection of an archaeal RNase P RNA mimics natural variation.
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-tRNA(Asp) 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.
authors.affiliation: Department of Microbiology, North Carolina State University, Raleigh, NC 27695-7615, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Williams
authors.forename: James W
authors.initials: JW
authors.lastname: Brown
mesh_terms: D001105:Archaea / Q000235:genetics*; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D019020:Directed Molecular Evolution; D014644:Genetic Variation; D017014:Methanobacterium / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D019642:RNA, Archaeal / Q000235:genetics*; D043262:Ribonuclease P / Q000235:genetics* / Q000378:metabolism
chemical_list: D019642:RNA, Archaeal; D043262:Ribonuclease P