{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T07:22:43Z","timestamp":1779348163711,"version":"3.51.4"},"reference-count":48,"publisher":"The Company of Biologists","issue":"7","license":[{"start":{"date-parts":[[2020,9,2]],"date-time":"2020-09-02T00:00:00Z","timestamp":1599004800000},"content-version":"vor","delay-in-days":2259,"URL":"http:\/\/creativecommons.org\/licenses\/by\/3.0"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2014,7,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n               <jats:p>The Anaphase-Promoting Complex (APC) is a multi-subunit E3 ubiquitin ligase that coordinates progression through the cell cycle by temporally and spatially promoting the degradation of key proteins. Many of these targeted proteins have been shown to play important roles in regulating orderly progression through the cell cycle. Using a previously described Drosophila in vitro expression cloning approach, we screened for new substrates of the APC in Xenopus egg extract and identified Drosophila MCPH1 (dMCPH1), a protein encoded by the homolog of a causative gene for autosomal recessive primary microcephaly in humans. The dMCPH1-B splice form, but not the dMCPH1-C splice form, undergoes robust degradation in Xenopus interphase egg extract in a Cdh1-dependent manner. Degradation of dMCPH1-B is controlled by an N-terminal destruction box (D-box) motif as its deletion or mutation blocks dMCPH1-B degradation. dMCPH1 levels are increased in Drosophila morula (APC2) mutant embryos, consistent with dMCPH1 being an APC substrate in vivo. Using a purified, reconstituted system, we show that dMCPH1-B is ubiquitinated by APCCdh1, indicating that the effect of APC on dMCPH1-B ubiquitination and degradation is direct. Full-length human MCPH1 (hMCPH1) has been predicted to be an APC substrate based on its interaction with the APC subunit Cdc27. We were not able to detect changes in hMCPH1 levels during the cell cycle in cultured human cells. Overexpression of hMCPH1 (or dMCPH1-B) in developing Xenopus embryos, however, disrupts cell division, suggesting that proper regulation of hMCPH1 and dMCPH1-B activity plays a critical role in proper cell-cycle progression.<\/jats:p>","DOI":"10.1242\/bio.20148318","type":"journal-article","created":{"date-parts":[[2014,6,28]],"date-time":"2014-06-28T05:11:39Z","timestamp":1403932299000},"page":"669-676","source":"Crossref","is-referenced-by-count":9,"title":["The <i>Drosophila<\/i> MCPH1-B isoform is a substrate of the APCCdh1 E3 ubiquitin ligase complex"],"prefix":"10.1242","volume":"3","author":[{"given":"Sarah G.","family":"Hainline","sequence":"first","affiliation":[{"name":"Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232-8240, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jamie L.","family":"Rickmyre","sequence":"additional","affiliation":[{"name":"Present address: Sarah Cannon Research Institute, Nashville, TN 37203, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Leif R.","family":"Neitzel","sequence":"additional","affiliation":[{"name":"Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232-8240, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laura A.","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232-8240, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ethan","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232-8240, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"237","published-online":{"date-parts":[[2014,6,27]]},"reference":[{"key":"2021111216404508200_b1","doi-asserted-by":"publisher","first-page":"725","DOI":"10.1038\/ncb1431","article-title":"Regulation of mitotic entry by microcephalin and its overlap with ATR signalling.","volume":"8","author":"Alderton","year":"2006","journal-title":"Nat. Cell Biol."},{"key":"2021111216404508200_b2","doi-asserted-by":"publisher","first-page":"2458","DOI":"10.1101\/gad.1361905","article-title":"A novel motif governs APC-dependent degradation of Drosophila ORC1 in vivo.","volume":"19","author":"Araki","year":"2005","journal-title":"Genes Dev."},{"key":"2021111216404508200_b3","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1016\/S0092-8674(03)00232-0","article-title":"Tome-1, a trigger of mitotic entry, is degraded during G1 via the APC.","volume":"113","author":"Ayad","year":"2003","journal-title":"Cell"},{"key":"2021111216404508200_b4","doi-asserted-by":"publisher","first-page":"74","DOI":"10.1038\/344074a0","article-title":"Cell cycle oscillation of phosphatase inhibitor-2 in rat fibroblasts coincident with p34cdc2 restriction.","volume":"344","author":"Brautigan","year":"1990","journal-title":"Nature"},{"key":"2021111216404508200_b5","doi-asserted-by":"publisher","first-page":"3578","DOI":"10.1242\/jcs.014290","article-title":"Microcephalin coordinates mitosis in the syncytial Drosophila embryo.","volume":"120","author":"Brunk","year":"2007","journal-title":"J. Cell Sci."},{"key":"2021111216404508200_b6","doi-asserted-by":"publisher","first-page":"2030","DOI":"10.4161\/cc.20643","article-title":"Regulation of E2F1 by APC\/C Cdh1 via K11 linkage-specific ubiquitin chain formation.","volume":"11","author":"Budhavarapu","year":"2012","journal-title":"Cell Cycle"},{"key":"2021111216404508200_b7","doi-asserted-by":"publisher","first-page":"1871","DOI":"10.1101\/gad.12.12.1871","article-title":"The checkpoint protein MAD2 and the mitotic regulator CDC20 form a ternary complex with the anaphase-promoting complex to control anaphase initiation.","volume":"12","author":"Fang","year":"1998","journal-title":"Genes Dev."},{"key":"2021111216404508200_b8","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1016\/S1097-2765(00)80126-4","article-title":"Direct binding of CDC20 protein family members activates the anaphase-promoting complex in mitosis and G1.","volume":"2","author":"Fang","year":"1998","journal-title":"Mol. Cell"},{"key":"2021111216404508200_b9","doi-asserted-by":"publisher","first-page":"411","DOI":"10.1016\/S0092-8674(00)00047-7","article-title":"The Xenopus chromokinesin Xkid is essential for metaphase chromosome alignment and must be degraded to allow anaphase chromosome movement.","volume":"102","author":"Funabiki","year":"2000","journal-title":"Cell"},{"key":"2021111216404508200_b10","doi-asserted-by":"publisher","first-page":"e40387","DOI":"10.1371\/journal.pone.0040387","article-title":"A novel MCPH1 isoform complements the defective chromosome condensation of human MCPH1-deficient cells.","volume":"7","author":"Gavvovidis","year":"2012","journal-title":"PLoS ONE"},{"key":"2021111216404508200_b11","doi-asserted-by":"publisher","first-page":"132","DOI":"10.1038\/349132a0","article-title":"Cyclin is degraded by the ubiquitin pathway.","volume":"349","author":"Glotzer","year":"1991","journal-title":"Nature"},{"key":"2021111216404508200_b12","volume-title":"Fly Pushing: The Theory and Practice of Drosophila Genetics","author":"Greenspan","year":"2004"},{"key":"2021111216404508200_b13","doi-asserted-by":"publisher","first-page":"1325","DOI":"10.1038\/ncb2342","article-title":"MCPH1 regulates the neuroprogenitor division mode by coupling the centrosomal cycle with mitotic entry through the Chk1-Cdc25 pathway.","volume":"13","author":"Gruber","year":"2011","journal-title":"Nat. Cell Biol."},{"key":"2021111216404508200_b14","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1016\/j.cub.2003.12.031","article-title":"The Drosophila MOS ortholog is not essential for meiosis.","volume":"14","author":"Ivanovska","year":"2004","journal-title":"Curr. Biol."},{"key":"2021111216404508200_b15","doi-asserted-by":"publisher","first-page":"136","DOI":"10.1086\/341283","article-title":"Identification of microcephalin, a protein implicated in determining the size of the human brain.","volume":"71","author":"Jackson","year":"2002","journal-title":"Am. J. Hum. Genet."},{"key":"2021111216404508200_b16","doi-asserted-by":"publisher","first-page":"279","DOI":"10.1016\/0092-8674(95)90338-0","article-title":"A 20S complex containing CDC27 and CDC16 catalyzes the mitosis-specific conjugation of ubiquitin to cyclin B. Cell","volume":"81","author":"King","year":"1995"},{"key":"2021111216404508200_b17","doi-asserted-by":"publisher","first-page":"1343","DOI":"10.1091\/mbc.7.9.1343","article-title":"Mutagenic analysis of the destruction signal of mitotic cyclins and structural characterization of ubiquitinated intermediates.","volume":"7","author":"King","year":"1996","journal-title":"Mol. Biol. Cell"},{"key":"2021111216404508200_b18","doi-asserted-by":"publisher","first-page":"973","DOI":"10.1126\/science.277.5328.973","article-title":"Expression cloning in the test tube.","volume":"277","author":"King","year":"1997","journal-title":"Science"},{"key":"2021111216404508200_b19","doi-asserted-by":"publisher","first-page":"2236","DOI":"10.1107\/S0907444913018593","article-title":"Building a pseudo-atomic model of the anaphase-promoting complex.","volume":"69","author":"Kulkarni","year":"2013","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"2021111216404508200_b20","doi-asserted-by":"publisher","first-page":"435","DOI":"10.1016\/j.devcel.2004.12.008","article-title":"Drosophila genome-scale screen for PAN GU kinase substrates identifies Mat89Bb as a cell cycle regulator.","volume":"8","author":"Lee","year":"2005","journal-title":"Dev. Cell"},{"key":"2021111216404508200_b21","doi-asserted-by":"publisher","first-page":"21393","DOI":"10.1074\/jbc.M110.208793","article-title":"SET nuclear oncogene associates with microcephalin\/MCPH1 and regulates chromosome condensation.","volume":"286","author":"Leung","year":"2011","journal-title":"J. Biol. Chem."},{"key":"2021111216404508200_b22","doi-asserted-by":"publisher","first-page":"15105","DOI":"10.1073\/pnas.0507722102","article-title":"BRIT1\/MCPH1 is a DNA damage responsive protein that regulates the Brca1-Chk1 pathway, implicating checkpoint dysfunction in microcephaly.","volume":"102","author":"Lin","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2021111216404508200_b23","doi-asserted-by":"publisher","first-page":"3565","DOI":"10.1093\/emboj\/17.13.3565","article-title":"Fizzy is required for activation of the APC\/cyclosome in Xenopus egg extracts.","volume":"17","author":"Lorca","year":"1998","journal-title":"EMBO J."},{"key":"2021111216404508200_b24","doi-asserted-by":"publisher","first-page":"6322","DOI":"10.1073\/pnas.1002447107","article-title":"A genome-scale protein interaction profile of Drosophila p53 uncovers additional nodes of the human p53 network.","volume":"107","author":"Lunardi","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2021111216404508200_b25","doi-asserted-by":"publisher","first-page":"1043","DOI":"10.1016\/S0092-8674(00)81209-X","article-title":"Geminin, an inhibitor of DNA replication, is degraded during mitosis.","volume":"93","author":"McGarry","year":"1998","journal-title":"Cell"},{"key":"2021111216404508200_b26","doi-asserted-by":"publisher","first-page":"482","DOI":"10.1016\/j.semcdb.2012.01.015","article-title":"Substrate targeting by the ubiquitin\u2013proteasome system in mitosis.","volume":"23","author":"Min","year":"2012","journal-title":"Semin. Cell Dev. Biol."},{"key":"2021111216404508200_b27","doi-asserted-by":"publisher","first-page":"3956","DOI":"10.4161\/cc.9.19.13162","article-title":"APC\/C(Cdc20) targets E2F1 for degradation in prometaphase.","volume":"9","author":"Peart","year":"2010","journal-title":"Cell Cycle"},{"key":"2021111216404508200_b28","doi-asserted-by":"publisher","first-page":"865","DOI":"10.1038\/ncb1895","article-title":"BRIT1\/MCPH1 links chromatin remodelling to DNA damage response.","volume":"11","author":"Peng","year":"2009","journal-title":"Nat. Cell Biol."},{"key":"2021111216404508200_b29","doi-asserted-by":"publisher","first-page":"644","DOI":"10.1038\/nrm1988","article-title":"The anaphase promoting complex\/cyclosome: a machine designed to destroy.","volume":"7","author":"Peters","year":"2006","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"2021111216404508200_b30","doi-asserted-by":"publisher","first-page":"655","DOI":"10.1101\/gad.14.6.655","article-title":"The KEN box: an APC recognition signal distinct from the D box targeted by Cdh1.","volume":"14","author":"Pfleger","year":"2000","journal-title":"Genes Dev."},{"key":"2021111216404508200_b31","doi-asserted-by":"publisher","first-page":"2396","DOI":"10.1101\/gad.918201","article-title":"Substrate recognition by the Cdc20 and Cdh1 components of the anaphase-promoting complex.","volume":"15","author":"Pfleger","year":"2001","journal-title":"Genes Dev."},{"key":"2021111216404508200_b32","doi-asserted-by":"publisher","first-page":"1759","DOI":"10.1101\/gad.897901","article-title":"Inhibition of Cdh1-APC by the MAD2-related protein MAD2L2: a novel mechanism for regulating Cdh1.","volume":"15","author":"Pfleger","year":"2001","journal-title":"Genes Dev."},{"key":"2021111216404508200_b33","doi-asserted-by":"publisher","first-page":"241","DOI":"10.1016\/j.gde.2005.04.009","article-title":"Evolution of primary microcephaly genes and the enlargement of primate brains.","volume":"15","author":"Ponting","year":"2005","journal-title":"Curr. Opin. Genet. Dev."},{"key":"2021111216404508200_b34","doi-asserted-by":"publisher","first-page":"1139","DOI":"10.1083\/jcb.200203035","article-title":"The roles of Fzy\/Cdc20 and Fzr\/Cdh1 in regulating the destruction of cyclin B in space and time.","volume":"157","author":"Raff","year":"2002","journal-title":"J. Cell Biol."},{"key":"2021111216404508200_b35","doi-asserted-by":"publisher","first-page":"145","DOI":"10.1016\/j.ccr.2006.07.002","article-title":"BRIT1 regulates early DNA damage response, chromosomal integrity, and cancer.","volume":"10","author":"Rai","year":"2006","journal-title":"Cancer Cell"},{"key":"2021111216404508200_b36","doi-asserted-by":"publisher","first-page":"185","DOI":"10.1016\/j.molcel.2005.03.017","article-title":"Sororin, a substrate of the anaphase-promoting complex, is required for sister chromatid cohesion in vertebrates.","volume":"18","author":"Rankin","year":"2005","journal-title":"Mol. Cell"},{"key":"2021111216404508200_b37","doi-asserted-by":"crossref","first-page":"3543","DOI":"10.1242\/dev.124.18.3543","article-title":"The Drosophila gene morula inhibits mitotic functions in the endo cell cycle and the mitotic cell cycle.","volume":"124","author":"Reed","year":"1997","journal-title":"Development"},{"key":"2021111216404508200_b38","doi-asserted-by":"publisher","first-page":"3565","DOI":"10.1242\/jcs.016626","article-title":"The Drosophila homolog of MCPH1, a human microcephaly gene, is required for genomic stability in the early embryo.","volume":"120","author":"Rickmyre","year":"2007","journal-title":"J. Cell Sci."},{"key":"2021111216404508200_b39","doi-asserted-by":"publisher","first-page":"2854","DOI":"10.1074\/jbc.M111.307868","article-title":"Molecular basis for the association of microcephalin (MCPH1) protein with the cell division cycle protein 27 (Cdc27) subunit of the anaphase-promoting complex.","volume":"287","author":"Singh","year":"2012","journal-title":"J. Biol. Chem."},{"key":"2021111216404508200_b40","doi-asserted-by":"publisher","first-page":"1294","DOI":"10.1101\/gr.269102","article-title":"The Drosophila gene collection: identification of putative full-length cDNAs for 70% of D. melanogaster genes.","volume":"12","author":"Stapleton","year":"2002","journal-title":"Genome Res."},{"key":"2021111216404508200_b41","doi-asserted-by":"publisher","first-page":"1149","DOI":"10.1083\/jcb.200810159","article-title":"Microcephalin and pericentrin regulate mitotic entry via centrosome-associated Chk1.","volume":"185","author":"Tibelius","year":"2009","journal-title":"J. Cell Biol."},{"key":"2021111216404508200_b42","doi-asserted-by":"publisher","first-page":"322","DOI":"10.4161\/cc.5.3.2412","article-title":"Misregulated chromosome condensation in MCPH1 primary microcephaly is mediated by condensin II.","volume":"5","author":"Trimborn","year":"2006","journal-title":"Cell Cycle"},{"key":"2021111216404508200_b43","doi-asserted-by":"publisher","first-page":"683","DOI":"10.1101\/gad.1071303","article-title":"Under arrest: cytostatic factor (CSF)-mediated metaphase arrest in vertebrate eggs.","volume":"17","author":"Tunquist","year":"2003","journal-title":"Genes Dev."},{"key":"2021111216404508200_b44","doi-asserted-by":"publisher","first-page":"194","DOI":"10.1038\/nature02381","article-title":"Degradation of the SCF component Skp2 in cell-cycle phase G1 by the anaphase-promoting complex.","volume":"428","author":"Wei","year":"2004","journal-title":"Nature"},{"key":"2021111216404508200_b45","doi-asserted-by":"publisher","first-page":"717","DOI":"10.1086\/429930","article-title":"Autosomal recessive primary microcephaly (MCPH): a review of clinical, molecular, and evolutionary findings.","volume":"76","author":"Woods","year":"2005","journal-title":"Am. J. Hum. Genet."},{"key":"2021111216404508200_b46","doi-asserted-by":"publisher","first-page":"841","DOI":"10.1083\/jcb.201106141","article-title":"MCPH1 regulates chromosome condensation and shaping as a composite modulator of condensin II.","volume":"194","author":"Yamashita","year":"2011","journal-title":"J. Cell Biol."},{"key":"2021111216404508200_b47","doi-asserted-by":"publisher","first-page":"907","DOI":"10.1038\/embor.2008.128","article-title":"MCPH1\/BRIT1 cooperates with E2F1 in the activation of checkpoint, DNA repair and apoptosis.","volume":"9","author":"Yang","year":"2008","journal-title":"EMBO Rep."},{"key":"2021111216404508200_b48","doi-asserted-by":"publisher","first-page":"418","DOI":"10.1126\/science.285.5426.418","article-title":"Identification of a vertebrate sister-chromatid separation inhibitor involved in transformation and tumorigenesis.","volume":"285","author":"Zou","year":"1999","journal-title":"Science"}],"container-title":["Biology Open"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.logists.com\/bio\/bio\/article-pdf\/3\/7\/669\/2114307\/bio-03-07-669.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/journals.logists.com\/bio\/bio\/article-pdf\/3\/7\/669\/2114307\/bio-03-07-669.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,12]],"date-time":"2021-11-12T16:41:04Z","timestamp":1636735264000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.biologists.com\/bio\/article\/3\/7\/669\/1115\/The-Drosophila-MCPH1-B-isoform-is-a-substrate-of"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,6,27]]},"references-count":48,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2014,7,15]]}},"URL":"https:\/\/doi.org\/10.1242\/bio.20148318","relation":{},"ISSN":["2046-6390"],"issn-type":[{"value":"2046-6390","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,6,27]]}}}