title: Spindle assembly checkpoint proteins are positioned close to core microtubule attachment sites at kinetochores.
abstract: Spindle assembly checkpoint proteins have been thought to reside in the peripheral corona region of the kinetochore, distal to microtubule attachment sites at the outer plate. However, recent biochemical evidence indicates that checkpoint proteins are closely linked to the core kinetochore microtubule attachment site comprised of the Knl1-Mis12-Ndc80 (KMN) complexes/KMN network. In this paper, we show that the Knl1-Zwint1 complex is required to recruit the Rod-Zwilch-Zw10 (RZZ) and Mad1-Mad2 complexes to the outer kinetochore. Consistent with this, nanometer-scale mapping indicates that RZZ, Mad1-Mad2, and the C terminus of the dynein recruitment factor Spindly are closely juxtaposed with the KMN network in metaphase cells when their dissociation is blocked and the checkpoint is active. In contrast, the N terminus of Spindly is ∼75 nm outside the calponin homology domain of the Ndc80 complex. These results reveal how checkpoint proteins are integrated within the substructure of the kinetochore and will aid in understanding the coordination of microtubule attachment and checkpoint signaling during chromosome segregation.
authors: Department of Biology, The University of North Carolina-Chapel Hill, Chapel Hill, NC 27599, USA. dileep@email.unc.edu
authors: Dileep
authors: D
authors: Varma
authors: Xiaohu
authors: X
authors: Wan
authors: Dhanya
authors: D
authors: Cheerambathur
authors: Reto
authors: R
authors: Gassmann
authors: Aussie
authors: A
authors: Suzuki
authors: Josh
authors: J
authors: Lawrimore
authors: Arshad
authors: A
authors: Desai
authors: E D
authors: ED
authors: Salmon
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D017173:Caenorhabditis elegans / Q000166:cytology / Q000378:metabolism; D004398:Dyneins / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000187:drug effects / Q000378:metabolism*; D059566:M Phase Cell Cycle Checkpoints / Q000187:drug effects; D008677:Metaphase / Q000187:drug effects; D008868:Microtubule Proteins / Q000737:chemistry / Q000378:metabolism; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism*; D050505:Mutant Proteins / Q000378:metabolism; D015739:Nocodazole / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000378:metabolism
chemical_list: D008868:Microtubule Proteins; D050505:Mutant Proteins; D021122:Protein Subunits; D049452:Green Fluorescent Proteins; D004398:Dyneins; D015739:Nocodazole
references: Mol Biol Cell. 2010 Jun 15;21(12):1968-81
references: 20427577
references: Cell. 2009 May 15;137(4):672-84
references: 19450515
references: J Cell Biol. 2009 Jun 1;185(5):859-74
references: 19468067
references: Mol Biol Cell. 2011 Mar 15;22(6):759-68
references: 21270439
references: Curr Biol. 2005 May 10;15(9):856-61
references: 15886105
references: J Cell Sci. 2012 Dec 15;125(Pt 24):5927-36
references: 23418356
references: Genes Dev. 2008 Sep 1;22(17):2385-99
references: 18765790
references: Philos Trans R Soc Lond B Biol Sci. 2011 Dec 27;366(1584):3595-604
references: 22084386
references: Nat Cell Biol. 2011 Apr;13(4):475-82
references: 21394085
references: Curr Biol. 2012 Nov 20;22(22):R966-80
references: 23174302
references: Mol Biol Cell. 2012 Mar;23(6):1035-46
references: 22298429
references: Mol Biol Cell. 2011 Sep;22(18):3318-30
references: 21775627
references: Mol Biol Cell. 2001 Jul;12(7):1995-2009
references: 11451998
references: Cell Mol Life Sci. 2010 Jul;67(13):2163-72
references: 20336345
references: J Cell Sci. 2000 Jun;113 ( Pt 11):1939-50
references: 10806105
references: J Biol Chem. 2004 Dec 24;279(52):54590-8
references: 15485811
references: Mol Biol Cell. 2008 Feb;19(2):587-94
references: 18045986
references: Curr Biol. 2004 Jun 8;14(11):953-64
references: 15182668
references: Genes Dev. 2010 May;24(9):957-71
references: 20439434
references: Curr Biol. 2007 Dec 18;17(24):2143-9
references: 18065224
references: Curr Biol. 2003 Dec 2;13(23):2103-9
references: 14654001
references: J Cell Biol. 2010 Sep 6;190(5):835-52
references: 20819937
references: Curr Biol. 2008 Nov 25;18(22):1778-84
references: 19026543
references: Nat Cell Biol. 2000 Dec;2(12):944-7
references: 11146660
references: Proc Natl Acad Sci U S A. 2012 Apr 24;109(17):6549-54
references: 22493223
references: Nat Cell Biol. 2001 Nov;3(11):1001-7
references: 11715021
references: J Cell Biol. 2008 Oct 20;183(2):187-94
references: 18936247
references: Trends Cell Biol. 2011 Mar;21(3):133-40
references: 21106376
references: J Cell Biol. 2008 Feb 11;180(3):507-20
references: 18268100
references: Methods Enzymol. 2003;360:597-617
references: 12622170
references: J Cell Biol. 2007 Jun 18;177(6):1005-15
references: 17576797
references: Trends Cell Biol. 2005 Jul;15(7):386-92
references: 15922598
references: J Cell Biol. 2005 Apr 11;169(1):49-60
references: 15824131
references: Oncogene. 2006 Nov 2;25(52):6901-14
references: 16732327
references: Dev Cell. 2007 Nov;13(5):663-676
references: 17981135
references: PLoS One. 2009 Oct 28;4(10):e7640
references: 19893618
references: J Cell Biol. 2000 Nov 13;151(4):739-48
references: 11076960
references: Structure. 2010 May 12;18(5):616-26
references: 20462495
references: Mol Biol Cell. 2005 Feb;16(2):519-31
references: 15548592
references: J Cell Sci. 2010 Mar 15;123(Pt 6):825-35
references: 20200228
references: Nat Rev Mol Cell Biol. 2007 May;8(5):379-93
references: 17426725
references: Curr Biol. 2004 Jan 6;14(1):56-61
references: 14711415
references: J Cell Biol. 2001 Dec 24;155(7):1159-72
references: 11756470