title: Evidence that substrate-specific effects of C5 protein lead to uniformity in binding and catalysis by RNase P.
abstract: The ribonucleoprotein enzyme RNase P processes all pre-tRNAs, yet some substrates apparently lack consensus elements for recognition. Here, we compare binding affinities and cleavage rates of Escherichia coli pre-tRNAs that exhibit the largest variation from consensus recognition sequences. These results reveal that the affinities of both consensus and nonconsensus substrates for the RNase P holoenzyme are essentially uniform. Comparative analyses of pre-tRNA and tRNA binding to the RNase P holoenzyme and P RNA alone reveal differential contributions of the protein subunit to 5' leader and tRNA affinity. Additionally, these studies reveal that uniform binding results from variations in the energetic contribution of the 5' leader, which serve to compensate for weaker tRNA interactions. Furthermore, kinetic analyses reveal uniformity in the rates of substrate cleavage that result from dramatic (> 900-fold) contributions of the protein subunit to catalysis for some nonconsensus pre-tRNAs. Together, these data suggest that an important biological function of RNase P protein is to offset differences in pre-tRNA structure such that binding and catalysis are uniform.
authors.affiliation: Center for RNA Molecular Biology and Department of Biochemistry, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
authors.forename: Lei
authors.initials: L
authors.lastname: Sun
authors.forename: Frank E
authors.initials: FE
authors.lastname: Campbell
authors.forename: Nathan H
authors.initials: NH
authors.lastname: Zahler
authors.forename: Michael E
authors.initials: ME
authors.lastname: Harris
mesh_terms: D024506:5' Flanking Region; D001483:Base Sequence; D001665:Binding Sites; D002384:Catalysis; D029968:Escherichia coli Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D012322:RNA Precursors / Q000737:chemistry; D012329:RNA, Bacterial / Q000737:chemistry; D012343:RNA, Transfer / Q000737:chemistry; D043262:Ribonuclease P / Q000737:chemistry*; D013379:Substrate Specificity
chemical_list: D029968:Escherichia coli Proteins; D012322:RNA Precursors; D012329:RNA, Bacterial; D012343:RNA, Transfer; D043262:Ribonuclease P; C470865:ribonuclease P, E coli