title: Identification of Endogenous mRNA-Binding Proteins in Yeast Using Crosslinking and PolyA Enrichment.
abstract: The maturation, localization, stability, and translation of messenger RNAs (mRNAs) are regulated by a wide variety of mRNA-binding proteins. Identification of the complete set of mRNA-binding proteins is a key step in understanding the regulation of gene expression. Herein, we describe a method for identifying yeast mRNA-binding proteins in a systematic manner using UV crosslinking, purification of polyA(+) mRNAs under denaturing conditions, and mass spectrometry to identify covalently bound proteins.
authors.affiliation: Department of Chemistry and Biochemistry, Howard Hughes Medical Institute, University of Colorado Boulder, Jennie Smoly Caruthers Biotech Bldg., 3415 Colorado Avenue, Boulder, CO, 80303, USA. sarah.f.mitchell@colorado.edu.
authors.forename: Sarah F
authors.initials: SF
authors.lastname: Mitchell
authors.affiliation: Department of Chemistry and Biochemistry, Howard Hughes Medical Institute, University of Colorado Boulder, Jennie Smoly Caruthers Biotech Bldg., 3415 Colorado Avenue, Boulder, CO, 80303, USA.
authors.forename: Roy
authors.initials: R
authors.lastname: Parker
mesh_terms: D005656:Fungal Proteins / Q000032:analysis / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods; D011061:Poly A / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000032:analysis / Q000378:metabolism*; D014466:Ultraviolet Rays; D015003:Yeasts / Q000737:chemistry / Q000254:growth & development / Q000378:metabolism*
chemical_list: D005656:Fungal Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011061:Poly A
keywords: Mass spectrometry; PolyA; Yeast; mRNA; mRNA-binding proteins