Eukaryotic elongation factor 2 kinase regulates the synthesis of microtubule-related proteins in neurons.
Modulation of the elongation phase of protein synthesis is important for numerous physiological processes in both neurons and other cell types. Elongation is primarily regulated via eukaryotic elongation factor 2 kinase (eEF2K). However, the consequence of altering eEF2K activity on the synthesis of specific proteins is largely unknown. Using both pharmacological and genetic manipulations of eEF2K combined with two protein-labeling techniques, stable isotope labeling of amino acids in cell culture and bio-orthogonal non-canonical amino acid tagging, we identified a subset of proteins whose synthesis is sensitive to inhibition of eEF2K in murine primary cortical neurons. Gene ontology (GO) analyses indicated that processes related to microtubules are particularly sensitive to eEF2K inhibition. Our findings suggest that eEF2K likely contributes to neuronal function by regulating the synthesis of microtubule-related proteins. Modulation of the elongation phase of protein synthesis is important for numerous physiological processes in neurons. Here, using labeling of new proteins coupled with proteomic techniques in primary cortical neurons, we find that the synthesis of microtubule-related proteins is up-regulated by inhibition of elongation. This suggests that translation elongation is a key regulator of cytoskeletal dynamics in neurons.
Centre for Biological Sciences, University of Southampton, Southampton, UK.
Justin W
JW
Kenney
Centre for Biological Sciences, University of Southampton, Southampton, UK.
Maja
M
Genheden
Centre for High-throughput Biology and Department of Biochemistry & Molecular Biology, University of British Columbia, Vancouver, BC, Canada.
Kyung-Mee
KM
Moon
Centre for Biological Sciences, University of Southampton, Southampton, UK.
Xuemin
X
Wang
Centre for High-throughput Biology and Department of Biochemistry & Molecular Biology, University of British Columbia, Vancouver, BC, Canada.
Leonard J
LJ
Foster
Centre for Biological Sciences, University of Southampton, Southampton, UK.
Christopher G
CG
Proud
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D054736:Elongation Factor 2 Kinase / Q000235:genetics / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D063990:Gene Ontology; D007554:Isotopes / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008868:Microtubule Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D052066:N-Ethylmaleimide-Sensitive Proteins / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D014176:Protein Biosynthesis; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology
D000596:Amino Acids; D004791:Enzyme Inhibitors; D007554:Isotopes; D008868:Microtubule Proteins; D054736:Elongation Factor 2 Kinase; D052066:N-Ethylmaleimide-Sensitive Proteins; C498733:Nsf protein, mouse
SILAC; elongation; mass spectrometry; microtubules; primary neurons; translational control