title: Substrate recognition and cleavage-site selection by a single-subunit protein-only RNase P.
abstract: RNase P is the enzyme that removes 5' extensions from tRNA precursors. With its diversity of enzyme forms-either protein- or RNA-based, ranging from single polypeptides to multi-subunit ribonucleoproteins-the RNase P enzyme family represents a unique model system to compare the evolution of enzymatic mechanisms. Here we present a comprehensive study of substrate recognition and cleavage-site selection by the nuclear single-subunit proteinaceous RNase P PRORP3 from Arabidopsis thaliana. Compared to bacterial RNase P, the best-characterized RNA-based enzyme form, PRORP3 requires a larger part of intact tRNA structure, but little to no determinants at the cleavage site or interactions with the 5' or 3' extensions of the tRNA. The cleavage site depends on the combined dimensions of acceptor stem and T domain, but also requires the leader to be single-stranded. Overall, the single-subunit PRORP appears mechanistically more similar to the complex nuclear ribonucleoprotein enzymes than to the simpler bacterial RNase P. Mechanistic similarity or dissimilarity among different forms of RNase P thus apparently do not necessarily reflect molecular composition or evolutionary relationship.
authors.affiliation: Center for Anatomy & Cell Biology, Medical University of Vienna, 1090 Vienna, Austria.
authors.forename: Nadia
authors.initials: N
authors.lastname: Brillante
authors.affiliation: Institute of Pharmaceutical Chemistry, Philipps-University Marburg, 35037 Marburg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Gößringer
authors.affiliation: Center for Anatomy & Cell Biology, Medical University of Vienna, 1090 Vienna, Austria.
authors.forename: Dominik
authors.initials: D
authors.lastname: Lindenhofer
authors.affiliation: Center for Anatomy & Cell Biology, Medical University of Vienna, 1090 Vienna, Austria.
authors.forename: Ursula
authors.initials: U
authors.lastname: Toth
authors.affiliation: Center for Anatomy & Cell Biology, Medical University of Vienna, 1090 Vienna, Austria walter.rossmanith@meduniwien.ac.at.
authors.forename: Walter
authors.initials: W
authors.lastname: Rossmanith
authors.affiliation: Institute of Pharmaceutical Chemistry, Philipps-University Marburg, 35037 Marburg, Germany roland.hartmann@staff.uni-marburg.de.
authors.forename: Roland K
authors.initials: RK
authors.lastname: Hartmann
mesh_terms: D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000235:genetics; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D019143:Evolution, Molecular; D015870:Gene Expression; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012322:RNA Precursors / Q000737:chemistry* / Q000378:metabolism; D012329:RNA, Bacterial / Q000737:chemistry / Q000378:metabolism; D018749:RNA, Plant / Q000737:chemistry* / Q000378:metabolism; D012343:RNA, Transfer / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D043262:Ribonuclease P / Q000737:chemistry* / Q000235:genetics; D013379:Substrate Specificity; D016963:Thermus thermophilus / Q000201:enzymology / Q000235:genetics
chemical_list: D029681:Arabidopsis Proteins; D007527:Isoenzymes; D012322:RNA Precursors; D012329:RNA, Bacterial; D018749:RNA, Plant; D011994:Recombinant Proteins; D012343:RNA, Transfer; C581753:PRORP1 protein, Arabidopsis; C000609578:PRORP3 protein, Arabidopsis; D043262:Ribonuclease P