title: A Suite of Biochemical Assays for Screening RNA Methyltransferase BCDIN3D.
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.
authors: 1 Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada.
authors: Levi L
authors: LL
authors: Blazer
authors: 1 Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada.
authors: Fengling
authors: F
authors: Li
authors: 1 Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada.
authors: Steven
authors: S
authors: Kennedy
authors: 2 Department of Pharmaceutical and Biomedical Sciences, University of Georgia, Athens, GA, USA.
authors: Yujun George
authors: YG
authors: Zheng
authors: 1 Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada.|3 Princess Margaret Cancer Centre and Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada.
authors: Cheryl H
authors: CH
authors: Arrowsmith
authors: 1 Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada.|4 Department of Pharmacology and Toxicology, University of Toronto, Toronto, ON, Canada.
authors: Masoud
authors: M
authors: Vedadi
mesh_terms: D001665:Binding Sites; D057075:Enzyme Assays / Q000379:methods*; D004795:Enzyme Stability; D005454:Fluorescence Polarization; D057166:High-Throughput Screening Assays; D006801:Humans; D007700:Kinetics; D008780:Methyltransferases / Q000378:metabolism*; D035683:MicroRNAs / Q000378:metabolism; D012313:RNA / Q000378:metabolism*; D012436:S-Adenosylmethionine; D020349:Surface Plasmon Resonance; D013696:Temperature
chemical_list: C541010:MIRN145 microRNA, human; D035683:MicroRNAs; D012313:RNA; D012436:S-Adenosylmethionine; C000622632:BCDIN3D protein, human; D008780:Methyltransferases
keywords: BCDIN3D; O-methyltransferase; RNA methyltransferase; microRNA