CK2 phosphorylation of eukaryotic translation initiation factor 5 potentiates cell cycle progression.

Department of Biomolecular Sciences, Fukushima Medical University School of Medicine, Fukushima 960-1295, Japan.
Proceedings of the National Academy of Sciences (Impact Factor: 9.81). 11/2005; 102(43):15688-93. DOI: 10.1073/pnas.0506791102
Source: PubMed

ABSTRACT 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.

  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The translation factor eIF5 is an important partner of eIF2, directly modulating its function in several critical steps. First, eIF5 binds eIF2/GTP/Met-tRNA(i)(Met) ternary complex (TC), promoting its recruitment to 40S ribosomal subunits. Secondly, its GTPase activating function promotes eIF2 dissociation for ribosomal subunit joining. Finally, eIF5 GDP dissociation inhibition (GDI) activity can antagonize eIF2 reactivation by competing with the eIF2 guanine exchange factor (GEF), eIF2B. The C-terminal domain (CTD) of eIF5, a W2-type HEAT domain, mediates its interaction with eIF2. Here, we characterize a related human protein containing MA3- and W2-type HEAT domains, previously termed BZW2 and renamed here as eIF5-mimic protein 1 (5MP1). Human 5MP1 interacts with eIF2 and eIF3 and inhibits general and gene-specific translation in mammalian systems. We further test whether 5MP1 is a mimic or competitor of the GEF catalytic subunit eIF2Bε or eIF5, using yeast as a model. Our results suggest that 5MP1 interacts with yeast eIF2 and promotes TC formation, but inhibits TC binding to the ribosome. Moreover, 5MP1 is not a GEF but a weak GDI for yeast eIF2. We propose that 5MP1 is a partial mimic and competitor of eIF5, interfering with the key steps by which eIF5 regulates eIF2 function.
    Nucleic Acids Research 07/2011; 39(19):8314-28. · 8.81 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Protein kinase CK2 (formerly known as casein kinase II) is a ubiquitious Ser/Thr kinase present in all eukaryotes. The α (catalytic) and β (regulatory) subunits of CK2 exist both as a tetrameric holoenzyme and as monomers in eukaryotic cells. CK2 has been implicated in multiple developmental and stress-responsive pathways including light signalling and circadian clock in plants. Recent studies using CK2 knockout and dominant negative mutants in Arabidopsis have uncovered new roles for this enzyme. CK2 substrates that have been identified so far are primarily transcription factors or regulatory proteins. CK2-mediated phosphorylation of these factors often results in alteration of the protein function including changes in the DNA-binding affinity, dimerization, stability, protein-protein interactions, and subcellular localization. CK2 has evolved as an essential housekeeping kinase in plants that modifies protein function in a dynamic way. This review summarizes the current knowledge of the role of CK2 in plant development.
    Journal of Experimental Botany 12/2013; · 5.79 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: mRNA translation is the most energy consuming process in the cell. In addition, it plays a pivotal role in the control of gene expression and is therefore tightly regulated. In response to various extracellular stimuli and intracellular cues, signaling pathways induce quantitative and qualitative changes in mRNA translation by modulating the phosphorylation status and thus the activity of components of the translational machinery. In this work we focus on the phosphoinositide 3-kinase (PI3K)/AKT and the mitogen-activated protein kinase (MAPK) pathways, as they are strongly implicated in the regulation of translation in homeostasis, whereas their malfunction has been linked to aberrant translation in human diseases, including cancer.
    Cold Spring Harbor perspectives in biology 08/2012; · 9.63 Impact Factor

Full-text (2 Sources)

Available from
Sep 2, 2014