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Structural perspective on the activation of RNAse P RNA by protein
Structural perspective on the activation of RNAse P RNA by protein.
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Ribonucleoprotein particles are central to numerous cellular pathways, but their study in vitro is often complicated by heterogeneity and aggregation. We describe a new technique to characterize these complexes trapped as homogeneous species in a nondenaturing gel. Using this technique, in conjunction with phosphorothioate footprinting analysis, we identify the protein-binding site and RNA folding states of ribonuclease P (RNase P), an RNA-based enzyme that, in vivo, requires a protein cofactor to catalyze the 5' maturation of precursor transfer RNA (pre-tRNA). Our results show that the protein binds to a patch of conserved RNA structure adjacent to the active site and influences the conformation of the RNA near the tRNA-binding site. The data are consistent with a role of the protein in substrate recognition and support a new model of the holoenzyme that is based on a recently solved crystal structure of RNase P RNA.
Buck AH, Kazantsev AV, Dalby AB, Pace NR
Nature structural & molecular biology
2005-11-01 00:00
12
11
958-64
Bacteria,Base Pairing,Base Sequence,DNA Footprinting,Electrophoretic Mobility Shift Assay,Evolution, Molecular,Models, Molecular,Molecular Sequence Data,Protein Binding,Protein Conformation,Protein Folding,RNA,Ribonuclease P,Structure-Activity Relationship,Substrate Specificity,RNA,Ribonuclease P
Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309, USA
Nat. Struct. Mol. Biol.
NIGMS GM-34527, NIGMS T32 GM-65103
1545-9993
10.1038/nsmb1004
nsmb1004
977
True
16228004
