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. 2003 Sep 1;374(Pt 2):349–358. doi: 10.1042/BJ20030600

Differential expression, localization and activity of two alternatively spliced isoforms of human APC regulator CDH1.

Yuan Zhou 1, Yick-Pang Ching 1, Raymond W M Ng 1, Dong-Yan Jin 1
PMCID: PMC1223613  PMID: 12797865

Abstract

The timely destruction of key regulators through ubiquitin-mediated proteolysis ensures the orderly progression of the cell cycle. The APC (anaphase-promoting complex) is a major component of this degradation machinery and its activation is required for the execution of critical events. Recent studies have just begun to reveal the complex control of the APC through a regulatory network involving WD40 repeat proteins CDC20 and CDH1. In the present paper, we report on the identification and characterization of human CDH1beta, a novel alternatively spliced isoform of CDH1. Both CDH1alpha and CDH1beta can bind to the APC and stimulate the degradation of cyclin B1, but they are differentially expressed in human tissues and cells. CDH1alpha contains a nuclear localization signal which is absent in CDH1beta. Intracellularly, CDH1alpha appears in the nucleus whereas CDH1beta is a predominantly cytoplasmic protein. The forced overexpression of CDH1alpha in cultured cells correlates with the reduction of nuclear cyclin A, but the steady-state amount of cyclin A does not change noticeably in CDH1beta-overexpressed cells. In Xenopus embryos, ectopic overexpression of human CDH1alpha, but not of CDH1beta, induces cell-cycle arrest during the first G(1) phase at the mid-blastula transition. Taken together, our findings document the differential expression, subcellular localization and cell-cycle-regulatory activity of human CDH1 isoforms.

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Selected References

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  1. Asakawa H., Kitamura K., Shimoda C. A novel Cdc20-related WD-repeat protein, Fzr1, is required for spore formation in Schizosaccharomyces pombe. Mol Genet Genomics. 2001 May;265(3):424–435. doi: 10.1007/s004380000429. [DOI] [PubMed] [Google Scholar]
  2. Bembenek J., Yu H. Regulation of the anaphase-promoting complex by the dual specificity phosphatase human Cdc14a. J Biol Chem. 2001 Oct 11;276(51):48237–48242. doi: 10.1074/jbc.M108126200. [DOI] [PubMed] [Google Scholar]
  3. Berry M. J., Banu L., Larsen P. R. Type I iodothyronine deiodinase is a selenocysteine-containing enzyme. Nature. 1991 Jan 31;349(6308):438–440. doi: 10.1038/349438a0. [DOI] [PubMed] [Google Scholar]
  4. Blanco M. A., Pelloquin L., Moreno S. Fission yeast mfr1 activates APC and coordinates meiotic nuclear division with sporulation. J Cell Sci. 2001 Jun;114(Pt 11):2135–2143. doi: 10.1242/jcs.114.11.2135. [DOI] [PubMed] [Google Scholar]
  5. Burton J. L., Solomon M. J. D box and KEN box motifs in budding yeast Hsl1p are required for APC-mediated degradation and direct binding to Cdc20p and Cdh1p. Genes Dev. 2001 Sep 15;15(18):2381–2395. doi: 10.1101/gad.917901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cartegni Luca, Chew Shern L., Krainer Adrian R. Listening to silence and understanding nonsense: exonic mutations that affect splicing. Nat Rev Genet. 2002 Apr;3(4):285–298. doi: 10.1038/nrg775. [DOI] [PubMed] [Google Scholar]
  7. Chen J., Fang G. MAD2B is an inhibitor of the anaphase-promoting complex. Genes Dev. 2001 Jul 15;15(14):1765–1770. doi: 10.1101/gad.898701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ching Yick-Pang, Zhou Hai-Jun, Yuan Jian-Gang, Qiang Bo-Qin, Kung Hf Hsiang-fu, Jin Dong-Yan. Identification and characterization of FTSJ2, a novel human nucleolar protein homologous to bacterial ribosomal RNA methyltransferase. Genomics. 2002 Jan;79(1):2–6. doi: 10.1006/geno.2001.6670. [DOI] [PubMed] [Google Scholar]
  9. Clute P., Pines J. Temporal and spatial control of cyclin B1 destruction in metaphase. Nat Cell Biol. 1999 Jun;1(2):82–87. doi: 10.1038/10049. [DOI] [PubMed] [Google Scholar]
  10. Cooper K. F., Mallory M. J., Egeland D. B., Jarnik M., Strich R. Ama1p is a meiosis-specific regulator of the anaphase promoting complex/cyclosome in yeast. Proc Natl Acad Sci U S A. 2000 Dec 19;97(26):14548–14553. doi: 10.1073/pnas.250351297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dawson I. A., Roth S., Artavanis-Tsakonas S. The Drosophila cell cycle gene fizzy is required for normal degradation of cyclins A and B during mitosis and has homology to the CDC20 gene of Saccharomyces cerevisiae. J Cell Biol. 1995 May;129(3):725–737. doi: 10.1083/jcb.129.3.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fang G., Yu H., Kirschner M. W. Direct binding of CDC20 protein family members activates the anaphase-promoting complex in mitosis and G1. Mol Cell. 1998 Aug;2(2):163–171. doi: 10.1016/s1097-2765(00)80126-4. [DOI] [PubMed] [Google Scholar]
  13. Fay David S., Keenan Sean, Han Min. fzr-1 and lin-35/Rb function redundantly to control cell proliferation in C. elegans as revealed by a nonbiased synthetic screen. Genes Dev. 2002 Feb 15;16(4):503–517. doi: 10.1101/gad.952302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gieffers C., Peters B. H., Kramer E. R., Dotti C. G., Peters J. M. Expression of the CDH1-associated form of the anaphase-promoting complex in postmitotic neurons. Proc Natl Acad Sci U S A. 1999 Sep 28;96(20):11317–11322. doi: 10.1073/pnas.96.20.11317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hagting Anja, Den Elzen Nicole, Vodermaier Hartmut C., Waizenegger Irene C., Peters Jan-Michael, Pines Jonathon. Human securin proteolysis is controlled by the spindle checkpoint and reveals when the APC/C switches from activation by Cdc20 to Cdh1. J Cell Biol. 2002 Jun 17;157(7):1125–1137. doi: 10.1083/jcb.200111001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Harper J. Wade, Burton Janet L., Solomon Mark J. The anaphase-promoting complex: it's not just for mitosis any more. Genes Dev. 2002 Sep 1;16(17):2179–2206. doi: 10.1101/gad.1013102. [DOI] [PubMed] [Google Scholar]
  17. Hilioti Z., Chung Y. S., Mochizuki Y., Hardy C. F., Cohen-Fix O. The anaphase inhibitor Pds1 binds to the APC/C-associated protein Cdc20 in a destruction box-dependent manner. Curr Biol. 2001 Sep 4;11(17):1347–1352. doi: 10.1016/s0960-9822(01)00399-2. [DOI] [PubMed] [Google Scholar]
  18. Hsu Jerry Y., Reimann Julie D. R., Sørensen Claus S., Lukas Jiri, Jackson Peter K. E2F-dependent accumulation of hEmi1 regulates S phase entry by inhibiting APC(Cdh1). Nat Cell Biol. 2002 May;4(5):358–366. doi: 10.1038/ncb785. [DOI] [PubMed] [Google Scholar]
  19. Inbal N., Listovsky T., Brandeis M. The mammalian Fizzy and Fizzy-related genes are regulated at the transcriptional and post-transcriptional levels. FEBS Lett. 1999 Dec 17;463(3):350–354. doi: 10.1016/s0014-5793(99)01640-3. [DOI] [PubMed] [Google Scholar]
  20. Jackman Mark, Kubota Yumiko, den Elzen Nicole, Hagting Anja, Pines Jonathon. Cyclin A- and cyclin E-Cdk complexes shuttle between the nucleus and the cytoplasm. Mol Biol Cell. 2002 Mar;13(3):1030–1045. doi: 10.1091/mbc.01-07-0361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jacobs Henning, Richter David, Venkatesh Tadmiri, Lehner Christian. Completion of mitosis requires neither fzr/rap nor fzr2, a male germline-specific Drosophila Cdh1 homolog. Curr Biol. 2002 Aug 20;12(16):1435–1441. doi: 10.1016/s0960-9822(02)01074-6. [DOI] [PubMed] [Google Scholar]
  22. Jaquenoud Malika, van Drogen Frank, Peter Matthias. Cell cycle-dependent nuclear export of Cdh1p may contribute to the inactivation of APC/C(Cdh1). EMBO J. 2002 Dec 2;21(23):6515–6526. doi: 10.1093/emboj/cdf634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jin D. Y., Chae H. Z., Rhee S. G., Jeang K. T. Regulatory role for a novel human thioredoxin peroxidase in NF-kappaB activation. J Biol Chem. 1997 Dec 5;272(49):30952–30961. doi: 10.1074/jbc.272.49.30952. [DOI] [PubMed] [Google Scholar]
  24. Jin D. Y., Giordano V., Kibler K. V., Nakano H., Jeang K. T. Role of adapter function in oncoprotein-mediated activation of NF-kappaB. Human T-cell leukemia virus type I Tax interacts directly with IkappaB kinase gamma. J Biol Chem. 1999 Jun 18;274(25):17402–17405. doi: 10.1074/jbc.274.25.17402. [DOI] [PubMed] [Google Scholar]
  25. Jin D. Y., Spencer F., Jeang K. T. Human T cell leukemia virus type 1 oncoprotein Tax targets the human mitotic checkpoint protein MAD1. Cell. 1998 Apr 3;93(1):81–91. doi: 10.1016/s0092-8674(00)81148-4. [DOI] [PubMed] [Google Scholar]
  26. Jin D. Y., Wang H. L., Zhou Y., Chun A. C., Kibler K. V., Hou Y. D., Kung H., Jeang K. T. Hepatitis C virus core protein-induced loss of LZIP function correlates with cellular transformation. EMBO J. 2000 Feb 15;19(4):729–740. doi: 10.1093/emboj/19.4.729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Jörgensen P. M., Brundell E., Starborg M., Hög C. A subunit of the anaphase-promoting complex is a centromere-associated protein in mammalian cells. Mol Cell Biol. 1998 Jan;18(1):468–476. doi: 10.1128/mcb.18.1.468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kotani S., Tanaka H., Yasuda H., Todokoro K. Regulation of APC activity by phosphorylation and regulatory factors. J Cell Biol. 1999 Aug 23;146(4):791–800. doi: 10.1083/jcb.146.4.791. [DOI] [PMC free article] [PubMed] [Google Scholar] [Retracted]
  29. Kramer E. R., Gieffers C., Hölzl G., Hengstschläger M., Peters J. M. Activation of the human anaphase-promoting complex by proteins of the CDC20/Fizzy family. Curr Biol. 1998 Nov 5;8(22):1207–1210. doi: 10.1016/s0960-9822(07)00510-6. [DOI] [PubMed] [Google Scholar]
  30. Kramer E. R., Scheuringer N., Podtelejnikov A. V., Mann M., Peters J. M. Mitotic regulation of the APC activator proteins CDC20 and CDH1. Mol Biol Cell. 2000 May;11(5):1555–1569. doi: 10.1091/mbc.11.5.1555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Littlepage Laurie E., Ruderman Joan V. Identification of a new APC/C recognition domain, the A box, which is required for the Cdh1-dependent destruction of the kinase Aurora-A during mitotic exit. Genes Dev. 2002 Sep 1;16(17):2274–2285. doi: 10.1101/gad.1007302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lorca T., Castro A., Martinez A. M., Vigneron S., Morin N., Sigrist S., Lehner C., Dorée M., Labbé J. C. Fizzy is required for activation of the APC/cyclosome in Xenopus egg extracts. EMBO J. 1998 Jul 1;17(13):3565–3575. doi: 10.1093/emboj/17.13.3565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Lukas C., Sørensen C. S., Kramer E., Santoni-Rugiu E., Lindeneg C., Peters J. M., Bartek J., Lukas J. Accumulation of cyclin B1 requires E2F and cyclin-A-dependent rearrangement of the anaphase-promoting complex. Nature. 1999 Oct 21;401(6755):815–818. doi: 10.1038/44611. [DOI] [PubMed] [Google Scholar]
  34. Modrek Barmak, Lee Christopher. A genomic view of alternative splicing. Nat Genet. 2002 Jan;30(1):13–19. doi: 10.1038/ng0102-13. [DOI] [PubMed] [Google Scholar]
  35. Musacchio Andrea, Hardwick Kevin G. The spindle checkpoint: structural insights into dynamic signalling. Nat Rev Mol Cell Biol. 2002 Oct;3(10):731–741. doi: 10.1038/nrm929. [DOI] [PubMed] [Google Scholar]
  36. Peters Jan-Michael. The anaphase-promoting complex: proteolysis in mitosis and beyond. Mol Cell. 2002 May;9(5):931–943. doi: 10.1016/s1097-2765(02)00540-3. [DOI] [PubMed] [Google Scholar]
  37. Pfleger C. M., Kirschner M. W. The KEN box: an APC recognition signal distinct from the D box targeted by Cdh1. Genes Dev. 2000 Mar 15;14(6):655–665. [PMC free article] [PubMed] [Google Scholar]
  38. Pfleger C. M., Salic A., Lee E., Kirschner M. W. Inhibition of Cdh1-APC by the MAD2-related protein MAD2L2: a novel mechanism for regulating Cdh1. Genes Dev. 2001 Jul 15;15(14):1759–1764. doi: 10.1101/gad.897901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Pines J. Four-dimensional control of the cell cycle. Nat Cell Biol. 1999 Jul;1(3):E73–E79. doi: 10.1038/11041. [DOI] [PubMed] [Google Scholar]
  40. Shirayama M., Tóth A., Gálová M., Nasmyth K. APC(Cdc20) promotes exit from mitosis by destroying the anaphase inhibitor Pds1 and cyclin Clb5. Nature. 1999 Nov 11;402(6758):203–207. doi: 10.1038/46080. [DOI] [PubMed] [Google Scholar]
  41. Sigrist S. J., Lehner C. F. Drosophila fizzy-related down-regulates mitotic cyclins and is required for cell proliferation arrest and entry into endocycles. Cell. 1997 Aug 22;90(4):671–681. doi: 10.1016/s0092-8674(00)80528-0. [DOI] [PubMed] [Google Scholar]
  42. Sorensen C. S., Lukas C., Kramer E. R., Peters J. M., Bartek J., Lukas J. Nonperiodic activity of the human anaphase-promoting complex-Cdh1 ubiquitin ligase results in continuous DNA synthesis uncoupled from mitosis. Mol Cell Biol. 2000 Oct;20(20):7613–7623. doi: 10.1128/mcb.20.20.7613-7623.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sørensen C. S., Lukas C., Kramer E. R., Peters J. M., Bartek J., Lukas J. A conserved cyclin-binding domain determines functional interplay between anaphase-promoting complex-Cdh1 and cyclin A-Cdk2 during cell cycle progression. Mol Cell Biol. 2001 Jun;21(11):3692–3703. doi: 10.1128/MCB.21.11.3692-3703.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Taylor S. S. Chromosome segregation: dual control ensures fidelity. 1999 Jul 29-Aug 12Curr Biol. 9(15):R562–R564. doi: 10.1016/s0960-9822(99)80355-8. [DOI] [PubMed] [Google Scholar]
  45. Visintin R., Prinz S., Amon A. CDC20 and CDH1: a family of substrate-specific activators of APC-dependent proteolysis. Science. 1997 Oct 17;278(5337):460–463. doi: 10.1126/science.278.5337.460. [DOI] [PubMed] [Google Scholar]
  46. Wan Y., Kirschner M. W. Identification of multiple CDH1 homologues in vertebrates conferring different substrate specificities. Proc Natl Acad Sci U S A. 2001 Oct 30;98(23):13066–13071. doi: 10.1073/pnas.231487598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wang C. X., Fisk B. C., Wadehra M., Su H., Braun J. Overexpression of murine fizzy-related (fzr) increases natural killer cell-mediated cell death and suppresses tumor growth. Blood. 2000 Jul 1;96(1):259–263. [PubMed] [Google Scholar]
  48. Wassmann K., Benezra R. Mitotic checkpoints: from yeast to cancer. Curr Opin Genet Dev. 2001 Feb;11(1):83–90. doi: 10.1016/s0959-437x(00)00161-1. [DOI] [PubMed] [Google Scholar]
  49. Weinstein J. Cell cycle-regulated expression, phosphorylation, and degradation of p55Cdc. A mammalian homolog of CDC20/Fizzy/slp1. J Biol Chem. 1997 Nov 7;272(45):28501–28511. doi: 10.1074/jbc.272.45.28501. [DOI] [PubMed] [Google Scholar]
  50. Weis Karsten. Regulating access to the genome: nucleocytoplasmic transport throughout the cell cycle. Cell. 2003 Feb 21;112(4):441–451. doi: 10.1016/s0092-8674(03)00082-5. [DOI] [PubMed] [Google Scholar]
  51. Yam C. H., Siu W. Y., Lau A., Poon R. Y. Degradation of cyclin A does not require its phosphorylation by CDC2 and cyclin-dependent kinase 2. J Biol Chem. 2000 Feb 4;275(5):3158–3167. doi: 10.1074/jbc.275.5.3158. [DOI] [PubMed] [Google Scholar]
  52. Yamaguchi S., Okayama H., Nurse P. Fission yeast Fizzy-related protein srw1p is a G(1)-specific promoter of mitotic cyclin B degradation. EMBO J. 2000 Aug 1;19(15):3968–3977. doi: 10.1093/emboj/19.15.3968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Yudkovsky Y., Shteinberg M., Listovsky T., Brandeis M., Hershko A. Phosphorylation of Cdc20/fizzy negatively regulates the mammalian cyclosome/APC in the mitotic checkpoint. Biochem Biophys Res Commun. 2000 May 10;271(2):299–304. doi: 10.1006/bbrc.2000.2622. [DOI] [PubMed] [Google Scholar]
  54. Zhou H. J., Wong C. M., Chen J. H., Qiang B. Q., Yuan J. G., Jin D. Y. Inhibition of LZIP-mediated transcription through direct interaction with a novel host cell factor-like protein. J Biol Chem. 2001 May 30;276(31):28933–28938. doi: 10.1074/jbc.M103893200. [DOI] [PubMed] [Google Scholar]
  55. Zhou Yuan, Ching Yick-Pang, Chun Abel C. S., Jin Dong-Yan. Nuclear localization of the cell cycle regulator CDH1 and its regulation by phosphorylation. J Biol Chem. 2003 Jan 29;278(14):12530–12536. doi: 10.1074/jbc.M212853200. [DOI] [PubMed] [Google Scholar]

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