Eukaryotic Elongation Factor 2 Kinase Activity Is Controlled by Multiple Inputs from Oncogenic Signaling

• Published in: Molecular and cellular biology Vol. 34; no. 22; pp. 4088 - 4103
• Main Authors: Wang, Xuemin, Regufe da Mota, Sergio, Liu, Rui, Moore, Claire E., Xie, Jianling, Lanucara, Francesco, Agarwala, Usha, Pyr dit Ruys, Sébastien, Vertommen, Didier, Rider, Mark H., Eyers, Claire E., Proud, Christopher G.
• Format: Journal Article
• Language: English
• Published: United States Taylor & Francis 01.11.2014; American Society for Microbiology
• Subjects: active sites; Animals; cell growth; Cell Line; Cell Line, Tumor; cell viability; Elongation Factor 2 Kinase - genetics; Elongation Factor 2 Kinase - metabolism; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases - metabolism; Humans; MAP Kinase Signaling System; Mechanistic Target of Rapamycin Complex 1; Mice; mitogen-activated protein kinase; Multiprotein Complexes - metabolism; Neoplasms - genetics; Neoplasms - metabolism; peptide elongation factors; Phosphorylation; Point Mutation; protein synthesis; raf Kinases - metabolism; ras Proteins - metabolism; Rats; Signal Transduction; starvation; tau-protein kinase; TOR Serine-Threonine Kinases - metabolism
• ISSN: 1098-5549; 0270-7306; 1098-5549
• DOI: 10.1128/MCB.01035-14

Eukaryotic elongation factor 2 kinase (eEF2K), an atypical calmodulin-dependent protein kinase, phosphorylates and inhibits eEF2, slowing down translation elongation. eEF2K contains an N-terminal catalytic domain, a C-terminal α-helical region and a linker containing several regulatory phosphorylation sites. eEF2K is expressed at high levels in certain cancers, where it may act to help cell survival, e.g., during nutrient starvation. However, it is a negative regulator of protein synthesis and thus cell growth, suggesting that cancer cells may possess mechanisms to inhibit eEF2K under good growth conditions, to allow protein synthesis to proceed. We show here that the mTORC1 pathway and the oncogenic Ras/Raf/MEK/extracellular signal-regulated kinase (ERK) pathway cooperate to restrict eEF2K activity. We identify multiple sites in eEF2K whose phosphorylation is regulated by mTORC1 and/or ERK, including new ones in the linker region. We demonstrate that certain sites are phosphorylated directly by mTOR or ERK. Our data reveal that glycogen synthase kinase 3 signaling also regulates eEF2 phosphorylation. In addition, we show that phosphorylation sites remote from the N-terminal calmodulin-binding motif regulate the phosphorylation of N-terminal sites that control CaM binding. Mutations in the former sites, which occur in cancer cells, cause the activation of eEF2K. eEF2K is thus regulated by a network of oncogenic signaling pathways.