Protein activation of a ribozyme: the role of bacterial RNase P protein.
Bacterial ribonuclease P (RNase P) belongs to a class of enzymes that utilize both RNAs and proteins to perform essential cellular functions. The bacterial RNase P protein is required to activate bacterial RNase P RNA in vivo, but previous studies have yielded contradictory conclusions regarding its specific functions. Here, we use biochemical and biophysical techniques to examine all of the proposed functions of the protein in both Escherichia coli and Bacillus subtilis RNase P. We demonstrate that the E. coli protein, but not the B. subtilis protein, stabilizes the global structure of RNase P RNA, although both proteins influence holoenzyme dimer formation and precursor tRNA recognition to different extents. By comparing each protein in complex with its cognate and noncognate RNA, we show that differences between the two types of holoenzymes reside primarily in the RNA and not the protein components of each. Our results reconcile previous contradictory conclusions regarding the role of the protein and support a model where the protein activates local RNA structures that manifest multiple holoenzyme properties.
Buck
Amy H
AH
Department of Chemistry and Biochemistry, University of Colorado at Boulder, Boulder, CO 80309, USA.
Dalby
Andrew B
AB
Poole
Alexander W
AW
Kazantsev
Alexei V
AV
Pace
Norman R
NR
D001412:Bacillus subtilis / Q000378:metabolism*; D004586:Electrophoresis; D024202:Electrophoretic Mobility Shift Assay; D004789:Enzyme Activation / Q000502:physiology; D004926:Escherichia coli / Q000378:metabolism*; D020035:Holoenzymes / Q000378:metabolism; D008954:Models, Biological; D016337:RNA, Catalytic / Q000378:metabolism*; D012343:RNA, Transfer / Q000378:metabolism*; D043262:Ribonuclease P / Q000235:genetics / Q000378:metabolism*; D013045:Species Specificity
D020035:Holoenzymes; D016337:RNA, Catalytic; D012343:RNA, Transfer; D043262:Ribonuclease P