Characterization of the aberrant splicing of MAP3K7 induced by cancer-associated SF3B1 mutation

J Biochem. 2021 Sep 22;170(1):69-77. doi: 10.1093/jb/mvab023.

Abstract

SF3B1, an essential RNA splicing factor, is frequently mutated in various types of cancers, and the cancer-associated SF3B1 mutation causes aberrant RNA splicing. The aberrant splicing of several transcripts, including MAP3K7, promotes tumorigenesis. Here, we identify a premature termination codon in the aberrantly spliced transcript of MAP3K7. Treatment of HEK293T cells transfected with the K700E-mutated SF3B1 with cycloheximide leads to increased accumulation of the aberrant spliced transcript of MAP3K7, demonstrating that the aberrantly spliced transcript of MAP3K7 is targeted by nonsense-mediated decay. The aberrantly spliced MAP3K7 transcript uses an aberrant 3' splice sites and an alternative branchpoint sequence. In addition, the aberrant splicing of MAP3K7 requires not only the polypyrimidine tract associated with normal splicing but also an alternative polypyrimidine tract upstream of the aberrant 3' splice site. Other cancer-associated SF3B1 mutations also cause the aberrant splicing of MAP3K7, which depends on the same sequence features. Our data provide a further understanding of the mechanisms underlying aberrant splicing induced by cancer-associated SF3B1 mutation, and reveal an important role of alternative polypyrimidine tract in diseases.

Keywords: 3ʹ; RNA splicing; branch point sequence; nonsense-mediated decay; polypyrimidine tract; splice site.

MeSH terms

  • Cells, Cultured
  • HEK293 Cells
  • Humans
  • MAP Kinase Kinase Kinases / genetics*
  • MAP Kinase Kinase Kinases / metabolism
  • Mutation
  • Neoplasms / genetics*
  • Neoplasms / metabolism
  • Phosphoproteins / genetics*
  • Phosphoproteins / metabolism
  • RNA Splicing / genetics*
  • RNA Splicing Factors / genetics*
  • RNA Splicing Factors / metabolism

Substances

  • Phosphoproteins
  • RNA Splicing Factors
  • SF3B1 protein, human
  • MAP Kinase Kinase Kinases
  • MAP kinase kinase kinase 7