Coupled Activation and Degradation of eEF2K Regulates Protein Synthesis in Response to Genotoxic Stress

Flore Kruiswijk, Laurensia Yuniati, Roberto Magliozzi, Teck Yew Low, Ratna Lim, Renske Bolder, Shabaz Mohammed, Christopher G. Proud, Albert J. R. Heck, Michele Pagano, Daniele Guardavaccaro*

*To whom correspondence should be addressed. E-mail: d.guardavaccaro{at}hubrecht.eu

This PDF file includes:

• Fig. S1. eEF2K interacts with βTrCP1 and βTrCP2.
• Fig. S2. Phosphorylation of the eEF2K degron in cells.
• Fig. S3. βTrCP-mediated ubiquitylation of eEF2K depends on an intact phosphodegron.
• Fig. S4. eEF2K abundance in response to mitogens and during the cell cycle.
• Fig. S5. The eEF2K degron is phosphorylated in response to genotoxic stress.
• Fig. S6. DNA damage–dependent phosphorylation of eEF2 on Thr⁵⁶ and decreased protein synthesis are mediated by eEF2K.
• Fig. S7. In vitro autophosphorylation of the eEF2K degron.
• Fig. S8. AMPK inhibition prevents eEF2K degradation during checkpoint silencing.
• Fig. S9. Serum starvation, a stimulus that activates AMPK, does not lead to eEF2K degradation.
• Fig. S10. The kinase-dead eEF2K mutant is not degraded during checkpoint silencing.
• Fig. S11. Expression of the eEF2K(S441A/S445A) mutant in exponentially growing U2OS cells does not lead to decreased protein synthesis.
  

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© 2012 American Association for the Advancement of Science