title: Nudel/NudE and Lis1 promote dynein and dynactin interaction in the context of spindle morphogenesis.
abstract: Lis1, Nudel/NudE, and dynactin are regulators of cytoplasmic dynein, a minus end-directed, microtubule (MT)-based motor required for proper spindle assembly and orientation. In vitro studies have shown that dynactin promotes processive movement of dynein on MTs, whereas Lis1 causes dynein to enter a persistent force-generating state (referred to here as dynein stall). Yet how the activities of Lis1, Nudel/NudE, and dynactin are coordinated to regulate dynein remains poorly understood in vivo. Working in Xenopus egg extracts, we show that Nudel/NudE facilitates the binding of Lis1 to dynein, which enhances the recruitment of dynactin to dynein. We further report a novel Lis1-dependent dynein-dynactin interaction that is essential for the organization of mitotic spindle poles. Finally, using assays for MT gliding and spindle assembly, we demonstrate an antagonistic relationship between Lis1 and dynactin that allows dynactin to relieve Lis1-induced dynein stall on MTs. Our findings suggest the interesting possibility that Lis1 and dynactin could alternately engage with dynein to allow the motor to promote spindle assembly.
authors.affiliation: Department of Embryology, Carnegie Institution for Science, Baltimore, MD 21218 Department of Biology, Johns Hopkins University, Baltimore, MD 21218 Department of Chemical Physiology, Scripps Research Institute, La Jolla, California 92037.
authors.forename: Shusheng
authors.initials: S
authors.lastname: Wang
authors.forename: Stephanie A
authors.initials: SA
authors.lastname: Ketcham
authors.forename: Arne
authors.initials: A
authors.lastname: Schön
authors.forename: Benjamin
authors.initials: B
authors.lastname: Goodman
authors.forename: Yueju
authors.initials: Y
authors.lastname: Wang
authors.forename: John
authors.initials: J
authors.lastname: Yates
authors.forename: Ernesto
authors.initials: E
authors.lastname: Freire
authors.forename: Trina A
authors.initials: TA
authors.lastname: Schroer
authors.forename: Yixian
authors.initials: Y
authors.lastname: Zheng
mesh_terms: D000818:Animals; D003598:Cytoskeletal Proteins; D000072159:Dynactin Complex; D004398:Dyneins / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D018507:Gene Expression Regulation, Developmental; D018386:Kinetochores / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D008869:Microtubule-Associated Proteins / Q000235:genetics* / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D009024:Morphogenesis / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D013094:Spermatozoa / Q000378:metabolism; D008941:Spindle Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D029867:Xenopus Proteins / Q000235:genetics* / Q000378:metabolism; D014982:Xenopus laevis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D015053:Zygote / Q000737:chemistry / Q000378:metabolism
chemical_list: D003598:Cytoskeletal Proteins; D000072159:Dynactin Complex; C558828:LIS1 protein, Xenopus; D008869:Microtubule-Associated Proteins; C555415:Ndel1 protein, Xenopus; D009687:Nuclear Proteins; D011994:Recombinant Proteins; D029867:Xenopus Proteins; D004398:Dyneins
references.citation: Neuron. 2004 Dec 2;44(5):809-21
references.pmid: 15572112
references.citation: Neuron. 2000 Dec;28(3):665-79
references.pmid: 11163258
references.citation: Nat Cell Biol. 2006 Jun;8(6):562-70
references.pmid: 16715075
references.citation: J Cell Biol. 2004 Feb 16;164(4):557-66
references.pmid: 14970193
references.citation: Methods Enzymol. 1998;298:171-84
references.pmid: 9751880
references.citation: Nat Cell Biol. 2012 Feb 29;14(3):224-30
references.pmid: 22373868
references.citation: Nat Genet. 2009 Feb;41(2):168-77
references.pmid: 19136950
references.citation: J Cell Sci. 2010 Jan 15;123(Pt 2):202-12
references.pmid: 20048338
references.citation: Anal Chem. 2004 Jul 15;76(14):4193-201
references.pmid: 15253663
references.citation: J Biol Chem. 2011 Nov 11;286(45):39615-22
references.pmid: 21911489
references.citation: Proc Natl Acad Sci U S A. 2009 Apr 7;106(14):5669-74
references.pmid: 19293377
references.citation: Nature. 2003 Feb 13;421(6924):715-8
references.pmid: 12610617
references.citation: Structure. 2012 Oct 10;20(10):1670-80
references.pmid: 22863569
references.citation: J Biol Chem. 1995 Dec 1;270(48):28806-11
references.pmid: 7499404
references.citation: Structure. 2007 Nov;15(11):1467-81
references.pmid: 17997972
references.citation: J Cell Biol. 1995 Dec;131(6 Pt 1):1507-16
references.pmid: 8522607
references.citation: J Cell Biol. 2011 Jun 27;193(7):1245-55
references.pmid: 21708978
references.citation: Nat Rev Mol Cell Biol. 2009 Dec;10(12):854-65
references.pmid: 19935668
references.citation: Nature. 2012 Mar 07;484(7394):345-50
references.pmid: 22398446
references.citation: J Neurosci. 2011 Nov 23;31(47):17207-19
references.pmid: 22114287
references.citation: J Biol Chem. 2012 Jul 20;287(30):24884-93
references.pmid: 22669947
references.citation: J Cell Biol. 1997 Sep 8;138(5):1055-66
references.pmid: 9281583
references.citation: J Cell Biol. 2011 Feb 7;192(3):433-45
references.pmid: 21282465
references.citation: Nat Cell Biol. 2000 Jan;2(1):20-4
references.pmid: 10620802
references.citation: Mol Biol Cell. 1995 Mar;6(3):297-310
references.pmid: 7612965
references.citation: J Cell Biol. 1991 Dec;115(5):1309-18
references.pmid: 1835460
references.citation: J Cell Biol. 2003 Feb 3;160(3):355-64
references.pmid: 12566428
references.citation: J Biol Chem. 2011 Jan 21;286(3):1959-65
references.pmid: 21036906
references.citation: Clin Genet. 2007 Oct;72(4):296-304
references.pmid: 17850624
references.citation: Neuron. 2000 Dec;28(3):697-711
references.pmid: 11163260
references.citation: J Cell Biol. 2011 Oct 17;195(2):193-201
references.pmid: 22006948
references.citation: Nature. 1993 Aug 19;364(6439):717-21
references.pmid: 8355785
references.citation: J Cell Biol. 2012 Jun 25;197(7):971-82
references.pmid: 22711696
references.citation: J Neurosci. 2006 Feb 15;26(7):2132-9
references.pmid: 16481446
references.citation: Nat Cell Biol. 2009 Mar;11(3):247-56
references.pmid: 19198602
references.citation: J Biol Chem. 2005 Jun 24;280(25):23960-5
references.pmid: 15826937
references.citation: Nat Struct Mol Biol. 2009 Mar;16(3):325-33
references.pmid: 19198589
references.citation: J Cell Biol. 2000 Aug 7;150(3):681-8
references.pmid: 10931877
references.citation: J Cell Biol. 2013 Apr 15;201(2):201-15
references.pmid: 23589491
references.citation: J Cell Biol. 2007 Aug 13;178(4):583-94
references.pmid: 17682047
references.citation: Cell. 2012 Aug 31;150(5):975-86
references.pmid: 22939623
references.citation: Mol Biol Cell. 2012 Oct;23(19):3827-37
references.pmid: 22918948
references.citation: Biochem Soc Trans. 2011 Oct;39(5):1169-78
references.pmid: 21936784
references.citation: Cell. 2010 Apr 16;141(2):304-14
references.pmid: 20403325
references.citation: J Biol Chem. 2011 Nov 11;286(45):39349-59
references.pmid: 21931160
references.citation: J Cell Biol. 1996 Oct;135(2):399-414
references.pmid: 8896597
references.citation: J Cell Biol. 2002 Mar 18;156(6):959-68
references.pmid: 11889140
references.citation: Annu Rev Cell Dev Biol. 2004;20:759-79
references.pmid: 15473859
references.citation: J Cell Biol. 2004 Aug 16;166(4):465-71
references.pmid: 15314063
references.citation: Mol Biol Cell. 2003 Dec;14(12):5089-97
references.pmid: 14565986
references.citation: Neuron. 2000 Dec;28(3):681-96
references.pmid: 11163259
references.citation: Rapid Commun Mass Spectrom. 2004;18(18):2162-8
references.pmid: 15317041
references.citation: J Biol Chem. 2011 Jan 7;286(1):587-93
references.pmid: 21056974
references.citation: Nature. 1996 Aug 1;382(6590):420-5
references.pmid: 8684481
references.citation: Science. 2011 Mar 4;331(6021):1159-65
references.pmid: 21330489
references.citation: Nat Cell Biol. 2000 Nov;2(11):767-75
references.pmid: 11056530
references.citation: J Cell Sci. 2010 Oct 15;123(Pt 20):3596-604
references.pmid: 20876661
references.citation: J Cell Biol. 1997 Aug 11;138(3):615-28
references.pmid: 9245790
references.citation: J Biol Chem. 2010 Nov 5;285(45):35238-44
references.pmid: 20826821
references.citation: Mol Biol Cell. 2003 Mar;14(3):871-88
references.pmid: 12631710
references.citation: Curr Biol. 2003 Mar 4;13(5):364-72
references.pmid: 12620184
references.citation: EMBO J. 2008 Oct 8;27(19):2471-83
references.pmid: 18784752