title: Phosphorylation of human eukaryotic initiation factor 2γ: novel site identification and targeted PKC involvement.
abstract: Eukaryotic translation requires a suite of proteins known as eukaryotic initiation factors (eIFs). These molecular effectors oversee the highly regulated initiation phase of translation. Essential to eukaryotic translation initiation is the protein eIF2, a heterotrimeric protein composed of the individually distinct subunits eIF2α, eIF2β, and eIF2γ. The ternary complex, formed when eIF2 binds to GTP and Met-tRNA(i), is responsible for shuttling Met-tRNA(i) onto the awaiting 40S ribosome. As a necessary component for translation initiation, much attention has been given to the phosphorylation of eIF2α. Despite several previous investigations into eIF2 phosphorylation, most have centered on α- or β-subunit phosphorylation and little is known regarding γ-subunit phosphorylation. Herein, we report eight sites of phosphorylation on the largest eIF2 subunit with seven novel phosphosite identifications via high resolution mass spectrometry. Of the eight sites identified, three are located in either the switch regions or nucleotide binding pocket domain. In addition, we have identified a possible kinase of eIF2, protein kinase C (PKC), which is capable of phosphorylating threonine 66 (thr-66) on the intact heterotrimer. These findings may shed new light on the regulation of ternary complex formation and alternate molecular effectors involved in this process prior to 80S ribosome formation and subsequent translation elongation and termination.
authors.affiliation: Department of Molecular and Cellular Biology and ‡Department of Biochemistry and Molecular Medicine, School of Medicine, University of California at Davis , Davis, California 95616, United States.
authors.forename: Armann
authors.initials: A
authors.lastname: Andaya
authors.forename: Weitao
authors.initials: W
authors.lastname: Jia
authors.forename: Masaaki
authors.initials: M
authors.lastname: Sokabe
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Fraser
authors.forename: John W B
authors.initials: JW
authors.lastname: Hershey
authors.forename: Julie A
authors.initials: JA
authors.lastname: Leary
mesh_terms: D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008954:Models, Biological; D010766:Phosphorylation; D011493:Protein Kinase C / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012343:RNA, Transfer / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D013912:Threonine / Q000737:chemistry
chemical_list: D015852:Eukaryotic Initiation Factor-2; D020543:Proteome; D013912:Threonine; D006160:Guanosine Triphosphate; D012343:RNA, Transfer; D011493:Protein Kinase C