title: Proximity mapping of human separase by the BioID approach.
abstract: Separase is a caspase-like cysteine protease that is best known for its essential role during the metaphase-to-anaphase transition when it cleaves the cohesin ring complex that keeps the sister chromatids together. Another important function of separase is to regulate the process of centriole separation, known as centriole disengagement, at the end of mitosis. We used proximity-dependent biotin identification (BioID) to expand our knowledge on the identity of separase's proximity interactors. We show that separase BioID labeled two domains at the mother centriole: an area underneath the centriolar appendages and another at the proximal end of the mother centriole. BioID analysis identified more than 200 proximity interactors of separase, one being the Alström Syndrome Protein 1 (ALMS1) at the base of centrioles. Other proximity interactors are the histone chaperons NAP1L1 and NAP1L4, which localize to the spindle poles during mitosis and the spindle assembly checkpoint proteins BUBR1, SKA1 and SKA3 that reside at kinetochores in early mitosis. Finally, we show that depletion of BUBR1 homolog from Caenorhabditis elegans delayed the recruitment of separase to mitotic chromosomes, and eventually anaphase onset.
authors: Zentrum für Molekulare Biologie der Universität Heidelberg, DKFZ-ZMBH Allianz, Im Neuenheimer Feld 282, 69120 Heidelberg, Germany.
authors: Fikret Gurkan
authors: FG
authors: Agircan
authors: Zentrum für Molekulare Biologie der Universität Heidelberg, DKFZ-ZMBH Allianz, Im Neuenheimer Feld 282, 69120 Heidelberg, Germany.
authors: Shoji
authors: S
authors: Hata
authors: Zentrum für Molekulare Biologie der Universität Heidelberg, DKFZ-ZMBH Allianz, Im Neuenheimer Feld 282, 69120 Heidelberg, Germany.
authors: Carmen
authors: C
authors: Nussbaum-Krammer
authors: Zentrum für Molekulare Biologie der Universität Heidelberg, DKFZ-ZMBH Allianz, Im Neuenheimer Feld 282, 69120 Heidelberg, Germany.
authors: Enrico
authors: E
authors: Atorino
authors: Zentrum für Molekulare Biologie der Universität Heidelberg, DKFZ-ZMBH Allianz, Im Neuenheimer Feld 282, 69120 Heidelberg, Germany. Electronic address: e.schiebel@zmbh.uni-heidelberg.de.
authors: Elmar
authors: E
authors: Schiebel
mesh_terms: D000705:Anaphase; D000818:Animals; D001681:Biological Assay; D001710:Biotin / Q000737:chemistry; D017173:Caenorhabditis elegans / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D018797:Cell Cycle Proteins; D045744:Cell Line, Tumor; D002502:Centrioles / Q000378:metabolism* / Q000648:ultrastructure; D018385:Centrosome / Q000378:metabolism* / Q000648:ultrastructure; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism; D002875:Chromosomes / Q000378:metabolism / Q000648:ultrastructure; D004625:Embryo, Nonmammalian; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D008677:Metaphase; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D056490:Nucleosome Assembly Protein 1 / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D064247:Separase / Q000235:genetics / Q000378:metabolism*
chemical_list: C490729:ALMS1 protein, human; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D008869:Microtubule-Associated Proteins; C533557:NAP1L1 protein, human; C104356:NAP1L4 protein, human; D009687:Nuclear Proteins; D056490:Nucleosome Assembly Protein 1; D011506:Proteins; C516427:SKA1 protein, human; C541140:Ska3 protein, human; D001710:Biotin; C584277:BUB1 protein, human; D017346:Protein Serine-Threonine Kinases; D064247:Separase
keywords: BioID; Centrosome; Kinetochore; Separase