title: Novel RNA-binding properties of Pop3p support a role for eukaryotic RNase P protein subunits in substrate recognition.
abstract: Ribonuclease P (RNase P) catalyzes the 5'-end maturation of transfer RNA molecules. Recent evidence suggests that the eukaryotic protein subunits may provide substrate-binding functions (True, H. L., and Celander, D. W. (1998) J. Biol. Chem. 273, 7193-7196). We now report that Pop3p, an essential protein subunit of the holoenzyme in Saccharomyces cerevisiae, displays novel RNA-binding properties. A recombinant form of Pop3p (H6Pop3p) displays a 3-fold greater affinity for binding pre-tRNA substrates relative to tRNA products. The recognition sequence for the H6Pop3p-substrate interaction in vitro was mapped to a 39-nucleotide long sequence that extends from position -21 to +18 surrounding the natural processing site in pre-tRNA substrates. H6Pop3p binds a variety of RNA molecules with high affinity (K(d) = 16-25 nm) and displays a preference for single-stranded RNAs. Removal or modification of basic C-terminal residues attenuates the RNA-binding properties displayed by the protein specifically for a pre-tRNA substrate. These studies support the model that eukaryotic RNase P proteins bind simultaneously to the RNA subunit and RNA substrate.
authors.affiliation: Department of Chemistry, Loyola University Chicago, Chicago, Illinois 60626, USA.
authors.forename: E M
authors.initials: EM
authors.lastname: Brusca
authors.forename: H L
authors.initials: HL
authors.lastname: True
authors.forename: D W
authors.initials: DW
authors.lastname: Celander
mesh_terms: D001665:Binding Sites; D004722:Endoribonucleases / Q000737:chemistry* / Q000378:metabolism; D021122:Protein Subunits; D012313:RNA / Q000378:metabolism*; D012322:RNA Precursors / Q000378:metabolism; D016337:RNA, Catalytic / Q000737:chemistry* / Q000378:metabolism; D043262:Ribonuclease P; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
chemical_list: D021122:Protein Subunits; D012322:RNA Precursors; D016337:RNA, Catalytic; D012313:RNA; D004722:Endoribonucleases; D043262:Ribonuclease P