Identification of human APC10/Doc1 as a subunit of anaphase promoting complex

Anaphase-promoting complex or cyclosome (APC) is a ubiquitin ligase which specifically targets mitotic regulatory factors such as Pds1/Cut2 and cyclin B. Identification of the subunits of multiprotein complex APC in several species revealed the highly conserved composition of APC from yeast to human. It has been reported, however, that vertebrate APC is composed of at least eight subunits, APC1 to APC8, while budding yeast APC is constituted of at least 12 components, Apc1 to Apc13. It has not yet been clearly understood whether additional components found in budding yeast, Apc9 to Apc13, are actually composed of mammalian APC. Here we isolated and characterized human APC10/Doc1, and found that APC10/Doc1 binds to APC core subunits throughout the cell cycle. Further, it was found that APC10/Doc1 is localized in centrosomes and mitotic spindles throughout mitosis, while it is also localized in kinetochores from prophase to anaphase and in midbody in telophase and cytokinesis. These results strongly support the notion that human APC10/Doc1 may be one of the APC core subunits rather than the transiently associated regulatory factor.

[1]  M. Starborg,et al.  A Subunit of the Anaphase-Promoting Complex Is a Centromere-Associated Protein in Mammalian Cells , 1998, Molecular and Cellular Biology.

[2]  E. Harlow,et al.  Antibodies: A Laboratory Manual , 1988 .

[3]  David O. Morgan,et al.  Regulation of the APC and the exit from mitosis , 1999, Nature Cell Biology.

[4]  M. Kirschner,et al.  Identification of a cullin homology region in a subunit of the anaphase-promoting complex. , 1998, Science.

[5]  J Weinstein,et al.  Cell Cycle-regulated Expression, Phosphorylation, and Degradation of p55Cdc , 1997, The Journal of Biological Chemistry.

[6]  Alexandre V. Podtelejnikov,et al.  Characterization of the DOC1/APC10 Subunit of the Yeast and the Human Anaphase-promoting Complex* , 1999, The Journal of Biological Chemistry.

[7]  E. Nigg,et al.  Cell Cycle Regulation of the Activity and Subcellular Localization of Plk1, a Human Protein Kinase Implicated in Mitotic Spindle Function Finally, Genetic Studies Performed with Fungi and Flies Have Identified Multiple Protein Kinases , 1995 .

[8]  K Nasmyth,et al.  Control of cyclin ubiquitination by CDK-regulated binding of Hct1 to the anaphase promoting complex. , 1998, Science.

[9]  M. Kirschner,et al.  Direct binding of CDC20 protein family members activates the anaphase-promoting complex in mitosis and G1. , 1998, Molecular cell.

[10]  A. Hershko,et al.  The cyclosome, a large complex containing cyclin-selective ubiquitin ligase activity, targets cyclins for destruction at the end of mitosis. , 1995, Molecular biology of the cell.

[11]  J. R. Daum,et al.  Mammalian p55CDC Mediates Association of the Spindle Checkpoint Protein Mad2 with the Cyclosome/Anaphase-promoting Complex, and is Involved in Regulating Anaphase Onset and Late Mitotic Events , 1998, The Journal of cell biology.

[12]  E. Nigg,et al.  The polo‐like kinase Plx1 is required for M phase exit and destruction of mitotic regulators in Xenopus egg extracts , 1998, The EMBO journal.

[13]  T. Toda,et al.  Requirement for PP1 phosphatase and 20S cyclosome/APC for the onset of anaphase is lessened by the dosage increase of a novel gene sds23+. , 1996, The EMBO journal.

[14]  C. Lehner,et al.  Fizzy is required for activation of the APC/cyclosome in Xenopus egg extracts , 1998, The EMBO journal.

[15]  P. Hieter,et al.  PKA and MPF-activated polo-like kinase regulate anaphase-promoting complex activity and mitosis progression. , 1998, Molecular cell.

[16]  S. Prinz,et al.  CDC20 and CDH1: a family of substrate-specific activators of APC-dependent proteolysis. , 1997, Science.

[17]  M. Kirschner,et al.  Identification of BIME as a Subunit of the Anaphase-Promoting Complex , 1996, Science.

[18]  M. Tyers,et al.  The phosphatase Cdc14 triggers mitotic exit by reversal of Cdk-dependent phosphorylation. , 1998, Molecular cell.

[19]  Angelika Amon,et al.  Cfi1 prevents premature exit from mitosis by anchoring Cdc14 phosphatase in the nucleolus , 1999, Nature.

[20]  A. Hershko,et al.  Reversible phosphorylation controls the activity of cyclosome-associated cyclin-ubiquitin ligase. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Stuart Tugendreich,et al.  CDC27Hs colocalizes with CDC16Hs to the centrosome and mitotic spindle and is essential for the metaphase to anaphase transition , 1995, Cell.

[22]  G. Wahl,et al.  PRC1: a human mitotic spindle-associated CDK substrate protein required for cytokinesis. , 1998, Molecular cell.

[23]  M. Kirschner,et al.  The checkpoint protein MAD2 and the mitotic regulator CDC20 form a ternary complex with the anaphase-promoting complex to control anaphase initiation. , 1998, Genes & development.

[24]  A. Murray,et al.  Budding yeast Cdc20: a target of the spindle checkpoint. , 1998, Science.

[25]  S. Elledge,et al.  How the Cyclin Became a Cyclin Regulated Proteolysis in the Cell Cycle , 1999, Cell.

[26]  A. Murray,et al.  Microinjection of Antibody to Mad2 Protein into Mammalian Cells in Mitosis Induces Premature Anaphase , 1998, The Journal of cell biology.

[27]  Tomohiro Matsumoto,et al.  Fission yeast Slp1: an effector of the Mad2-dependent spindle checkpoint. , 1998, Science.

[28]  Giovanna Lucchini,et al.  Budding Yeast Bub2 Is Localized at Spindle Pole Bodies and Activates the Mitotic Checkpoint via a Different Pathway from Mad2 , 1999, The Journal of cell biology.

[29]  S. Sazer,et al.  The Schizosaccharomyces pombe spindle checkpoint protein mad2p blocks anaphase and genetically interacts with the anaphase-promoting complex. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[30]  R. Benezra,et al.  MAD2 associates with the cyclosome/anaphase-promoting complex and inhibits its activity. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[31]  A. Murray,et al.  A novel yeast screen for mitotic arrest mutants identifies DOC1, a new gene involved in cyclin proteolysis. , 1997, Molecular biology of the cell.

[32]  Kim Nasmyth,et al.  The Polo‐like kinase Cdc5p and the WD‐repeat protein Cdc20p/fizzy are regulators and substrates of the anaphase promoting complex in Saccharomyces cerevisiae , 1998, The EMBO journal.

[33]  M. Kirschner,et al.  A 20s complex containing CDC27 and CDC16 catalyzes the mitosis-specific conjugation of ubiquitin to cyclin B , 1995, Cell.

[34]  Anna Shevchenko,et al.  Exit from Mitosis Is Triggered by Tem1-Dependent Release of the Protein Phosphatase Cdc14 from Nucleolar RENT Complex , 1999, Cell.

[35]  T. Toda,et al.  Apc10 and Ste9/Srw1, two regulators of the APC–cyclosome, as well as the CDK inhibitor Rum1 are required for G1 cell‐cycle arrest in fission yeast , 1998, The EMBO journal.

[36]  Hiroshi Yamada,et al.  20S cyclosome complex formation and proteolytic activity inhibited by the cAMP/PKA pathway , 1996, Nature.

[37]  K. Todokoro,et al.  Thrombopoietin-induced Polyploidization of Bone Marrow Megakaryocytes Is Due to a Unique Regulatory Mechanism in Late Mitosis , 1997, The Journal of cell biology.

[38]  K Nasmyth,et al.  Identification of Subunits of the Anaphase-Promoting Complex of Saccharomyces cerevisiae , 1996, Science.

[39]  D. Fesquet,et al.  A Bub2p‐dependent spindle checkpoint pathway regulates the Dbf2p kinase in budding yeast , 1999, The EMBO journal.

[40]  K Nasmyth,et al.  Sister chromatid separation and chromosome re‐duplication are regulated by different mechanisms in response to spindle damage , 1999, The EMBO journal.

[41]  David O. Morgan,et al.  The Polo-related kinase Cdc5 activates and is destroyed by the mitotic cyclin destruction machinery in S. cerevisiae , 1998, Current Biology.

[42]  Michael Schwab,et al.  Yeast Hct1 Is a Regulator of Clb2 Cyclin Proteolysis , 1997, Cell.

[43]  Jan-Michael Peters,et al.  Activation of the human anaphase-promoting complex by proteins of the CDC20/Fizzy family , 1998, Current Biology.

[44]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[45]  Tim Hunt,et al.  Cut2 proteolysis required for sister-chromatid separation in fission yeast , 1996, Nature.

[46]  Kim Nasmyth,et al.  Genes involved in sister chromatid separation are needed for b-type cyclin proteolysis in budding yeast , 1995, Cell.