title: GAK is phosphorylated by c-Src and translocated from the centrosome to chromatin at the end of telophase.
abstract: Cyclin G-associated kinase (GAK) harbors a consensus phosphorylation motif (Y412) for c-Src; however, its physiological significance remains elusive. Here, we show that GAK is phosphorylated by c-Src not only at Y412 but also at Y1149. An anti-GAK-pY412 antibody recognized the shifted band of GAK during M phase. Immunofluorescence (IF) showed that GAK-pY412/pY1149 signals were present in the nucleus during interphase, translocated to chromosomes at prophase and prometaphase, moved to centrosomes at metaphase, and finally translocated to chromosomes at the end of telophase, when nuclear membrane formation was almost complete. These subcellular movements of GAK resemble those of DNA licensing factors. Indeed, mass spectrometry identified mini-chromosome maintenance (MCM) 3, an essential component of the DNA licensing system, as one of the association partners of GAK; immunoprecipitation-mediated Western blotting confirmed their association in vivo. These results suggest that the c-Src_GAK_MCM axis plays an important role in cell cycle progression through control of the DNA replication licensing system.
authors.affiliation: a Department of Molecular Genetics , Research Institute for Microbial Diseases, Osaka University , Yamadaoka, Suita, Osaka , Japan.
authors.forename: Kohshiro
authors.initials: K
authors.lastname: Fukushima
authors.affiliation: a Department of Molecular Genetics , Research Institute for Microbial Diseases, Osaka University , Yamadaoka, Suita, Osaka , Japan.
authors.forename: Mian
authors.initials: M
authors.lastname: Wang
authors.affiliation: a Department of Molecular Genetics , Research Institute for Microbial Diseases, Osaka University , Yamadaoka, Suita, Osaka , Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Naito
authors.affiliation: b First Department of Oral and Maxillofacial Surgery , Graduate School of Dentistry, Osaka University , Yamadaoka, Suita, Osaka , Japan.
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Uchihashi
authors.affiliation: a Department of Molecular Genetics , Research Institute for Microbial Diseases, Osaka University , Yamadaoka, Suita, Osaka , Japan.
authors.forename: Yorika
authors.initials: Y
authors.lastname: Kato
authors.affiliation: a Department of Molecular Genetics , Research Institute for Microbial Diseases, Osaka University , Yamadaoka, Suita, Osaka , Japan.
authors.forename: Satomi
authors.initials: S
authors.lastname: Mukai
authors.affiliation: a Department of Molecular Genetics , Research Institute for Microbial Diseases, Osaka University , Yamadaoka, Suita, Osaka , Japan.
authors.forename: Norikazu
authors.initials: N
authors.lastname: Yabuta
authors.affiliation: a Department of Molecular Genetics , Research Institute for Microbial Diseases, Osaka University , Yamadaoka, Suita, Osaka , Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nojima
mesh_terms: D000906:Antibodies / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation; D018385:Centrosome / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007399:Interphase; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D064111:Minichromosome Maintenance Complex Component 2 / Q000378:metabolism; D064126:Minichromosome Maintenance Complex Component 3 / Q000378:metabolism; D008938:Mitosis; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D021381:Protein Transport; D016196:S Phase; D013347:Subcellular Fractions / Q000201:enzymology; D013692:Telophase; D019061:src-Family Kinases / Q000378:metabolism*
chemical_list: D000906:Antibodies; D002843:Chromatin; D047908:Intracellular Signaling Peptides and Proteins; C066470:MCM3 protein, human; D019061:src-Family Kinases; C486325:GAK protein, human; D017346:Protein Serine-Threonine Kinases; C084859:MCM2 protein, human; D064111:Minichromosome Maintenance Complex Component 2; D064126:Minichromosome Maintenance Complex Component 3
keywords: DNA replication licensing; GAK; MCM3; c-Src; phosphorylation