Downregulation of splicing factor SRSF3 induces p53β, an alternatively spliced isoform of p53 that promotes cellular senescence

Y. Tang, I. Horikawa, M. Ajiro, A. I. Robles, K. Fujita, A. M. Mondal, J. K. Stauffer, Z. M. Zheng, C. C. Harris

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

Most human pre-mRNA transcripts are alternatively spliced, but the significance and fine-tuning of alternative splicing in different biological processes is only starting to be understood. SRSF3 (SRp20) is a member of a highly conserved family of splicing factors that have critical roles in key biological processes, including tumor progression. Here, we show that SRSF3 regulates cellular senescence, a p53-mediated process to suppress tumorigenesis, through TP53 alternative splicing. Downregulation of SRSF3 was observed in normal human fibroblasts undergoing replicative senescence, and was associated with the upregulation of p53β, an alternatively spliced isoform of p53 that promotes p53-mediated senescence. Knockdown of SRSF3 by short interfering RNA (siRNA) in early-passage fibroblasts induced senescence, which was associated with elevated expression of p53β at mRNA and protein levels. Knockdown of p53 partially rescued SRSF3-knockdown-induced senescence, suggesting that SRSF3 acts on p53-mediated cellular senescence. RNA pulldown assays demonstrated that SRSF3 binds to an alternatively spliced exon uniquely included in p53β mRNA through the consensus SRSF3-binding sequences. RNA crosslinking and immunoprecipitation assays (CLIP) also showed that SRSF3 in vivo binds to endogenous p53 pre-mRNA at the region containing the p53β-unique exon. Splicing assays using a transfected TP53 minigene in combination with siRNA knockdown of SRSF3 showed that SRSF3 functions to inhibit the inclusion of the p53β-unique exon in splicing of p53 pre-mRNA. These data suggest that downregulation of SRSF3 represents an endogenous mechanism for cellular senescence that directly regulates the TP53 alternative splicing to generate p53β. This study uncovers the role for general splicing machinery in tumorigenesis, and suggests that SRSF3 is a direct regulator of p53.

Original languageEnglish (US)
Pages (from-to)2792-2798
Number of pages7
JournalOncogene
Volume32
Issue number22
DOIs
StatePublished - May 30 2013
Externally publishedYes

Fingerprint

Cell Aging
RNA Precursors
Protein Isoforms
Alternative Splicing
Down-Regulation
Biological Phenomena
Exons
Small Interfering RNA
Carcinogenesis
Fibroblasts
RNA
Messenger RNA
Immunoprecipitation
Up-Regulation
RNA Splicing Factors
Neoplasms
Proteins

Keywords

  • alternative splicing
  • cellular senescence
  • isoform
  • p53
  • SRSF3

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Tang, Y., Horikawa, I., Ajiro, M., Robles, A. I., Fujita, K., Mondal, A. M., ... Harris, C. C. (2013). Downregulation of splicing factor SRSF3 induces p53β, an alternatively spliced isoform of p53 that promotes cellular senescence. Oncogene, 32(22), 2792-2798. https://doi.org/10.1038/onc.2012.288

Downregulation of splicing factor SRSF3 induces p53β, an alternatively spliced isoform of p53 that promotes cellular senescence. / Tang, Y.; Horikawa, I.; Ajiro, M.; Robles, A. I.; Fujita, K.; Mondal, A. M.; Stauffer, J. K.; Zheng, Z. M.; Harris, C. C.

In: Oncogene, Vol. 32, No. 22, 30.05.2013, p. 2792-2798.

Research output: Contribution to journalArticle

Tang, Y, Horikawa, I, Ajiro, M, Robles, AI, Fujita, K, Mondal, AM, Stauffer, JK, Zheng, ZM & Harris, CC 2013, 'Downregulation of splicing factor SRSF3 induces p53β, an alternatively spliced isoform of p53 that promotes cellular senescence', Oncogene, vol. 32, no. 22, pp. 2792-2798. https://doi.org/10.1038/onc.2012.288
Tang, Y. ; Horikawa, I. ; Ajiro, M. ; Robles, A. I. ; Fujita, K. ; Mondal, A. M. ; Stauffer, J. K. ; Zheng, Z. M. ; Harris, C. C. / Downregulation of splicing factor SRSF3 induces p53β, an alternatively spliced isoform of p53 that promotes cellular senescence. In: Oncogene. 2013 ; Vol. 32, No. 22. pp. 2792-2798.
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