The RNA-binding protein Sam68 modulates the alternative splicing of Bcl-x

Maria Paola Paronetto, Tilman Achsel, Autumn Massiello, Charles E. Chalfant, Claudio Sette

Research output: Contribution to journalArticle

Abstract

The RNA-binding protein Sam68 is involved in apoptosis, but its cellular mRNA targets and its mechanism of action remain unknown. We demonstrate that Sam68 binds the mRNA for Bcl-x and affects its alternative splicing. Depletion of Sam68 by RNA interference caused accumulation of antiapoptotic Bcl-x(L), whereas its up-regulation increased the levels of proapoptotic Bcl-x(s). Tyrosine phosphorylation of Sam68 by Fyn inverted this effect and favored the Bcl-x(L) splice site selection. A point mutation in the RNA-binding domain of Sam68 influenced its splicing activity and subnuclear localization. Moreover, co-expression of ASF/SF2 with Sam68, or fusion with an RS domain, counteracted Sam68 splicing activity toward Bcl-x. Finally, Sam68 interacted with heterogenous nuclear RNP (hnRNP) A1, and depletion of hnRNP A1 or mutations that impair this interaction attenuated Bcl-x(s) splicing. Our results indicate that Sam68 plays a role in the regulation of Bcl-x alternative splicing and that tyrosine phosphorylation of Sam68 by Src-like kinases can switch its role from proapoptotic to antiapoptotic in live cells.

Original languageEnglish
Pages (from-to)929-939
Number of pages11
JournalJournal of Cell Biology
Volume176
Issue number7
DOIs
Publication statusPublished - Mar 26 2007

    Fingerprint

ASJC Scopus subject areas

  • Cell Biology

Cite this