The conserved serine-threonine kinase, Cdc7, plays a crucial role in initiation of DNA replication by facilitating the assembly of an initiation complex. Cdc7 is expressed at a high level and exhibits significant kinase activity not only during S-phase but also during G2/M-phases. A conserved mitotic kinase, Aurora B, is activated during M-phase by association with INCENP, forming the chromosome passenger complex with Borealin and Survivin. We show that Cdc7 phosphorylates and stimulates Aurora B kinase activity in vitro. We identified threonine-236 as a critical phosphorylation site on Aurora B that could be a target of Cdc7 or could be an autophosphorylation site stimulated by Cdc7-mediated phosphorylation elsewhere. We found that threonines at both 232 (that has been identified as an autophosphorylation site) and 236 are essential for the kinase activity of Aurora B. Cdc7 down regulation or inhibition reduced Aurora B activity in vivo and led to retarded M-phase progression. SAC imposed by paclitaxel was dramatically reversed by Cdc7 inhibition, similar to the effect of Aurora B inhibition under the similar situation. Our data show that Cdc7 contributes to M-phase progression and to spindle assembly checkpoint most likely through Aurora B activation.

Carna Biosciences Inc Kobe Japan

Danish Cancer Society Research Center Copenhagen Denmark

Department of Genome Medicine Tokyo Metropolitan Institute of Medical Science Tokyo 156 8506 Japan

Department of Molecular Oncology Institute of Development Aging and Cancer Tohoku University Sendai 980 8575 Japan

Department of Neural Regeneration and Cell Communication Mie University Graduate School of Medicine Tsu Mie 514 8507 Japan

Department of Physiology Mie University Graduate School of Medicine Tsu Mie 514 8507 Japan

Division of Genome Biology Department of Medical Biochemistry and Biophysics Karolinska Institute Stockholm Sweden

Institute of Molecular and Translational Medicine Faculty of Medicine and Dentistry Palacky University 77900 Olomouc Czech Republic

Laboratory of Protein Analyses Tokyo Metropolitan Institute of Medical Science Tokyo 156 8506 Japan

Medical Education Center Graduate School of Medicine Kyoto University Kyoto Japan

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