A Suite of Biochemical Assays for Screening RNA Methyltransferase BCDIN3D

SLAS Discov. 2017 Jan;22(1):32-39. doi: 10.1177/1087057116666276. Epub 2016 Sep 27.

Abstract

BCDIN3D is an RNA-methyltransferase that O-methylates the 5' phosphate of RNA and regulates microRNA maturation. To discover small-molecule inhibitors of BCDIN3D, a suite of biochemical assays was developed. A radiometric methyltransferase assay and fluorescence polarization-based S-adenosylmethionine and RNA displacement assays are described. In addition, differential scanning fluorimetry and surface plasmon resonance were used to characterize binding. These assays provide a comprehensive package for the development of small-molecule modulators of BCDIN3D activity.

Keywords: BCDIN3D; O-methyltransferase; RNA methyltransferase; microRNA.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Enzyme Assays / methods*
  • Enzyme Stability
  • Fluorescence Polarization
  • High-Throughput Screening Assays
  • Humans
  • Kinetics
  • Methyltransferases / metabolism*
  • MicroRNAs / metabolism
  • RNA / metabolism*
  • S-Adenosylmethionine
  • Surface Plasmon Resonance
  • Temperature

Substances

  • MIRN145 microRNA, human
  • MicroRNAs
  • RNA
  • S-Adenosylmethionine
  • BCDIN3D protein, human
  • Methyltransferases