Global analysis reveals SRp20- and SRp75-specific mRNPs in cycling and neural cells

Nat Struct Mol Biol. 2010 Aug;17(8):962-70. doi: 10.1038/nsmb.1862. Epub 2010 Jul 18.

Abstract

Members of the SR protein family of RNA-binding proteins have numerous roles in mRNA metabolism, from transcription to translation. To understand how SR proteins coordinate gene regulation, comprehensive knowledge of endogenous mRNA targets is needed. Here we establish physiological expression of GFP-tagged SR proteins from stable transgenes. Using the GFP tag for immunopurification of mRNPs, mRNA targets of SRp20 and SRp75 were identified in cycling and neurally induced P19 cells. Genome-wide analysis showed that SRp20 and SRp75 associate with hundreds of distinct, functionally related groups of transcripts that change in response to neural differentiation. Knockdown of either SRp20 or SRp75 led to up- or downregulation of specific transcripts, including identified targets, and rescue by the GFP-tagged SR proteins proved their functionality. Thus, SR proteins contribute to the execution of gene-expression programs through their association with distinct endogenous mRNAs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Cycle*
  • Cell Differentiation / genetics
  • Cell Line, Tumor
  • Chromosomes, Artificial, Bacterial / metabolism
  • Gene Expression Regulation, Neoplastic
  • Gene Knockdown Techniques
  • Green Fluorescent Proteins / metabolism
  • Immunoprecipitation
  • Mice
  • Neurons / cytology*
  • Neurons / metabolism*
  • Organ Specificity
  • Protein Binding
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism*
  • Ribonucleoproteins / metabolism*
  • Serine-Arginine Splicing Factors

Substances

  • RNA, Messenger
  • RNA-Binding Proteins
  • Ribonucleoproteins
  • SRp75 protein, mouse
  • Srsf3 protein, mouse
  • messenger ribonucleoprotein
  • Green Fluorescent Proteins
  • Serine-Arginine Splicing Factors