Format

Send to

Choose Destination
ACS Chem Biol. 2016 Aug 19;11(8):2285-92. doi: 10.1021/acschembio.6b00275. Epub 2016 Jun 23.

Determination of the Specificity Landscape for Ribonuclease P Processing of Precursor tRNA 5' Leader Sequences.

Author information

1
Department of Biochemistry, Case Western Reserve University School of Medicine , Cleveland, Ohio 44106, United States.
2
School of Business, CUNY Baruch College , New York, New York 10010, United States.
3
Center for RNA Molecular Biology, Case Western Reserve University School of Medicine , Cleveland, Ohio 44106, United States.

Abstract

Maturation of tRNA depends on a single endonuclease, ribonuclease P (RNase P), to remove highly variable 5' leader sequences from precursor tRNA transcripts. Here, we use high-throughput enzymology to report multiple-turnover and single-turnover kinetics for Escherichia coli RNase P processing of all possible 5' leader sequences, including nucleotides contacting both the RNA and protein subunits of RNase P. The results reveal that the identity of N(-2) and N(-3) relative to the cleavage site at N(1) primarily control alternative substrate selection and act at the level of association not the cleavage step. As a consequence, the specificity for N(-1), which contacts the active site and contributes to catalysis, is suppressed. This study demonstrates high-throughput RNA enzymology as a means to globally determine RNA specificity landscapes and reveals the mechanism of substrate discrimination by a widespread and essential RNA-processing enzyme.

PMID:
27336323
DOI:
10.1021/acschembio.6b00275
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for American Chemical Society
Loading ...
Support Center