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Coupled activation and degradation of eEF2K regulates protein synthesis in response to genotoxic stress

Coupled activation and degradation of eEF2K regulates protein synthesis in response to genotoxic stress
Coupled activation and degradation of eEF2K regulates protein synthesis in response to genotoxic stress
The kinase eEF2K [eukaryotic elongation factor 2 (eEF2) kinase] controls the rate of peptide chain elongation by phosphorylating eEF2, the protein that mediates the movement of the ribosome along the mRNA by promoting translocation of the transfer RNA from the A to the P site in the ribosome. eEF2K-mediated phosphorylation of eEF2 on threonine 56 (Thr??) decreases its affinity for the ribosome, thereby inhibiting elongation. Here, we show that in response to genotoxic stress, eEF2K was activated by AMPK (adenosine monophosphate-activated protein kinase)-mediated phosphorylation on serine 398. Activated eEF2K phosphorylated eEF2 and induced a temporary ribosomal slowdown at the stage of elongation. Subsequently, during DNA damage checkpoint silencing, a process required to allow cell cycle reentry, eEF2K was degraded by the ubiquitin-proteasome system through the ubiquitin ligase SCF(?TrCP) (Skp1-Cul1-F-box protein, ?-transducin repeat-containing protein) to enable rapid resumption of translation elongation. This event required autophosphorylation of eEF2K on a canonical ?TrCP-binding domain. The inability to degrade eEF2K during checkpoint silencing caused sustained phosphorylation of eEF2 on Thr?? and delayed the resumption of translation elongation. Our study therefore establishes a link between DNA damage signaling and translation elongation.
ra40
Kruiswijk, Flore
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Yuniati, Laurensia
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Magliozzi, Roberto
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Low, Teck Yew
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Lim, Ratna
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Bolder, Renske
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Mohammed, Shabaz
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Proud, Christopher G.
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Heck, Albert J.R.
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Pagano, Michele
f471d619-bc39-4893-9503-bfb1190bd6e8
Guardavaccaro, Daniele
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Kruiswijk, Flore
ee4fd0ed-9c30-4ee6-84e8-a458f9d06ad6
Yuniati, Laurensia
7a704881-310f-4018-bc90-3321bd1ce7f5
Magliozzi, Roberto
e046b0e1-9930-4a76-a61f-3222e0da9c0c
Low, Teck Yew
76d09a49-8770-4d2d-9f97-25640c268837
Lim, Ratna
b6019295-5723-412e-a655-c3023ad253c4
Bolder, Renske
72273752-c2ff-44ee-9fdc-55969eebb87b
Mohammed, Shabaz
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Proud, Christopher G.
59dabfc8-4b44-4be8-a17f-578a58550cb3
Heck, Albert J.R.
08ab14d3-9adc-46df-b22f-0d99ec58c171
Pagano, Michele
f471d619-bc39-4893-9503-bfb1190bd6e8
Guardavaccaro, Daniele
4ffb4197-4b8e-497d-b311-39d9502150fc

Kruiswijk, Flore, Yuniati, Laurensia, Magliozzi, Roberto, Low, Teck Yew, Lim, Ratna, Bolder, Renske, Mohammed, Shabaz, Proud, Christopher G., Heck, Albert J.R., Pagano, Michele and Guardavaccaro, Daniele (2012) Coupled activation and degradation of eEF2K regulates protein synthesis in response to genotoxic stress. Science Signaling, 5 (227), ra40. (doi:10.1126/scisignal.2002718). (PMID:22669845)

Record type: Article

Abstract

The kinase eEF2K [eukaryotic elongation factor 2 (eEF2) kinase] controls the rate of peptide chain elongation by phosphorylating eEF2, the protein that mediates the movement of the ribosome along the mRNA by promoting translocation of the transfer RNA from the A to the P site in the ribosome. eEF2K-mediated phosphorylation of eEF2 on threonine 56 (Thr??) decreases its affinity for the ribosome, thereby inhibiting elongation. Here, we show that in response to genotoxic stress, eEF2K was activated by AMPK (adenosine monophosphate-activated protein kinase)-mediated phosphorylation on serine 398. Activated eEF2K phosphorylated eEF2 and induced a temporary ribosomal slowdown at the stage of elongation. Subsequently, during DNA damage checkpoint silencing, a process required to allow cell cycle reentry, eEF2K was degraded by the ubiquitin-proteasome system through the ubiquitin ligase SCF(?TrCP) (Skp1-Cul1-F-box protein, ?-transducin repeat-containing protein) to enable rapid resumption of translation elongation. This event required autophosphorylation of eEF2K on a canonical ?TrCP-binding domain. The inability to degrade eEF2K during checkpoint silencing caused sustained phosphorylation of eEF2 on Thr?? and delayed the resumption of translation elongation. Our study therefore establishes a link between DNA damage signaling and translation elongation.

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Published date: 5 June 2012
Organisations: Centre for Biological Sciences

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Local EPrints ID: 350245
URI: http://eprints.soton.ac.uk/id/eprint/350245
PURE UUID: b7ef1695-8323-4b6e-9669-456dd105e921

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Date deposited: 20 Mar 2013 12:16
Last modified: 14 Mar 2024 13:23

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Contributors

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

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