title: The P4 metal binding site in RNase P RNA affects active site metal affinity through substrate positioning.
abstract: Although helix P4 in the catalytic domain of the RNase P ribozyme is known to coordinate magnesium ions important for activity, distinguishing between direct and indirect roles in catalysis has been difficult. Here, we provide evidence for an indirect role in catalysis by showing that while the universally conserved bulge of helix P4 is positioned 5 nt downstream of the cleavage site, changes in its structure can still purturb active site metal binding. Because changes in helix P4 also appear to alter its position relative to the pre-tRNA cleavage site, these data suggest that P4 contributes to catalytic metal ion binding through substrate positioning.
authors.affiliation: Center for RNA Molecular Biology, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA. elc3@case.edu
authors.forename: Eric L
authors.initials: EL
authors.lastname: Christian
authors.forename: Kari M J
authors.initials: KM
authors.lastname: Smith
authors.forename: Nicholas
authors.initials: N
authors.lastname: Perera
authors.forename: Michael E
authors.initials: ME
authors.lastname: Harris
mesh_terms: D001665:Binding Sites; D002104:Cadmium / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002384:Catalysis; D020134:Catalytic Domain; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D008670:Metals / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D009154:Mutation; D009690:Nucleic Acid Conformation; D010710:Phosphates / Q000378:metabolism; D012322:RNA Precursors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016337:RNA, Catalytic / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D043262:Ribonuclease P / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D008670:Metals; D010710:Phosphates; D012322:RNA Precursors; D016337:RNA, Catalytic; D002104:Cadmium; D043262:Ribonuclease P; D008274:Magnesium; D002118:Calcium