Tyrosine Phosphorylation of Caspase-8 Abrogates Its Apoptotic Activity and Promotes Activation of c-Src

PLoS One. 2016 Apr 21;11(4):e0153946. doi: 10.1371/journal.pone.0153946. eCollection 2016.

Abstract

Src family tyrosine kinases (SFKs) phosphorylate caspase-8A at tyrosine (Y) 397 resulting in suppression of apoptosis. In addition, the phosphorylation of caspase-8A at other sites including Y465 has been implicated in the regulation of caspase-8 activity. However, the functional consequences of these modifications on caspase-8 processing/activity have not been elucidated. Moreover, various Src substrates are known to act as potent Src regulators, but no such role has been explored for caspase-8. We asked whether the newly identified caspase-8 phosphorylation sites might regulate caspase-8 activation and conversely, whether caspase-8 phosphorylation might affect Src activity. Here we show that Src phosphorylates caspase-8A at multiple tyrosine sites; of these, we have focused on Y397 within the linker region and Y465 within the p12 subunit of caspase-8A. We show that phosphomimetic mutation of caspase-8A at Y465 prevents its cleavage and the subsequent activation of caspase-3 and suppresses apoptosis. Furthermore, simultaneous phosphomimetic mutation of caspase-8A at Y397 and Y465 promotes the phosphorylation of c-Src at Y416 and increases c-Src activity. Finally, we demonstrate that caspase-8 activity prevents its own tyrosine phosphorylation by Src. Together these data reveal that dual phosphorylation converts caspase-8 from a pro-apoptotic to a pro-survival mediator. Specifically, tyrosine phosphorylation by Src renders caspase-8 uncleavable and thereby inactive, and at the same time converts it to a Src activator. This novel dynamic interplay between Src and caspase-8 likely acts as a potent signal-integrating switch directing the cell towards apoptosis or survival.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Apoptosis*
  • Caspase 8 / chemistry
  • Caspase 8 / metabolism*
  • Cell Line
  • Enzyme Activation
  • Humans
  • Phosphorylation
  • Tyrosine / metabolism*
  • src-Family Kinases / metabolism*

Substances

  • Tyrosine
  • src-Family Kinases
  • Caspase 8

Grants and funding

This work was supported by Canadian Institutes of Health Research (http://www.cihr-irsc.gc.ca/e/193.html, grant number: MOP129493 to JM), and McMaster Medicine Internal Career Research Award (http://fhs.mcmaster.ca/medicine/research_awards.htm, E.J. Moran Campbell Internal Career Research Award to JT).