CK2 phosphorylation of eukaryotic translation initiation factor 5 potentiates cell cycle progression.
Casein kinase 2 (CK2) is a ubiquitous eukaryotic Ser/Thr protein kinase that plays an important role in cell cycle progression. Although its function in this process remains unclear, it is known to be required for the G(1) and G(2)/M phase transitions in yeast. Here, we show that CK2 activity changes notably during cell cycle progression and is increased within 3 h of serum stimulation of quiescent cells. During the time period in which it exhibits high enzymatic activity, CK2 associates with and phosphorylates a key molecule for translation initiation, eukaryotic translation initiation factor (eIF) 5. Using MS, we show that Ser-389 and -390 of eIF5 are major sites of phosphorylation by CK2. This is confirmed using eIF5 mutants that lack CK2 sites; the phosphorylation levels of mutant eIF5 proteins are significantly reduced, relative to WT eIF5, both in vitro and in vivo. Expression of these mutants reveals that they have a dominant-negative effect on phosphorylation of endogenous eIF5, and that they perturb synchronous progression of cells through S to M phase, resulting in a significant reduction in growth rate. Furthermore, the formation of mature eIF5/eIF2/eIF3 complex is reduced in these cells, and, in fact, restricted diffusional motion of WT eIF5 was almost abolished in a GFP-tagged eIF5 mutant lacking CK2 phosphorylation sites, as measured by fluorescence correlation spectroscopy. These results suggest that CK2 may be involved in the regulation of cell cycle progression by associating with and phosphorylating a key molecule for translation initiation.
Homma
Miwako Kato
MK
Department of Biomolecular Sciences, Fukushima Medical University School of Medicine, Fukushima 960-1295, Japan. mkhomma@fmu.ac.jp
Wada
Ikuo
I
Suzuki
Toshiyuki
T
Yamaki
Junko
J
Krebs
Edwin G
EG
Homma
Yoshimi
Y
D000818:Animals; D019556:COS Cells; D047390:Casein Kinase II / Q000502:physiology*; D002453:Cell Cycle; D002522:Chlorocebus aethiops; D040061:Eukaryotic Initiation Factor-5 / Q000378:metabolism*; D006801:Humans; D010766:Phosphorylation; D014176:Protein Biosynthesis; D013050:Spectrometry, Fluorescence
D040061:Eukaryotic Initiation Factor-5; D047390:Casein Kinase II