Arginine methylation controls the subcellular localization and functions of the oncoprotein splicing factor SF2/ASF

Mol Cell Biol. 2010 Jun;30(11):2762-74. doi: 10.1128/MCB.01270-09. Epub 2010 Mar 22.

Abstract

Alternative splicing and posttranslational modifications (PTMs) are major sources of protein diversity in eukaryotic proteomes. The SR protein SF2/ASF is an oncoprotein that functions in pre-mRNA splicing, with additional roles in other posttranscriptional and translational events. Functional studies of SR protein PTMs have focused exclusively on the reversible phosphorylation of Ser residues in the C-terminal RS domain. We confirmed that human SF2/ASF is methylated at residues R93, R97, and R109, which were identified in a global proteomic analysis of Arg methylation, and further investigated whether these methylated residues regulate the properties of SF2/ASF. We show that the three arginines additively control the subcellular localization of SF2/ASF and that both the positive charge and the methylation state are important. Mutations that block methylation and remove the positive charge result in the cytoplasmic accumulation of SF2/ASF. The consequent decrease in nuclear SF2/ASF levels prevents it from modulating the alternative splicing of target genes, results in higher translation stimulation, and abrogates the enhancement of nonsense-mediated mRNA decay. This study addresses the mechanisms by which Arg methylation and the associated positive charge regulate the activities of SF2/ASF and emphasizes the significance of localization control for an oncoprotein with multiple functions in different cellular compartments.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Alternative Splicing*
  • Amino Acid Sequence
  • Arginine / metabolism*
  • HeLa Cells
  • Humans
  • Methylation
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Oncogene Proteins / genetics
  • Oncogene Proteins / metabolism*
  • RNA Precursors / genetics
  • RNA Precursors / metabolism
  • RNA-Binding Proteins
  • Serine-Arginine Splicing Factors
  • Subcellular Fractions / metabolism
  • Tandem Mass Spectrometry

Substances

  • Nuclear Proteins
  • Oncogene Proteins
  • RNA Precursors
  • RNA-Binding Proteins
  • Serine-Arginine Splicing Factors
  • Arginine