title: Bicaudal D2, dynein, and kinesin-1 associate with nuclear pore complexes and regulate centrosome and nuclear positioning during mitotic entry.
abstract: BICD2 is one of the two mammalian homologues of the Drosophila Bicaudal D, an evolutionarily conserved adaptor between microtubule motors and their cargo that was previously shown to link vesicles and mRNP complexes to the dynein motor. Here, we identified a G2-specific role for BICD2 in the relative positioning of the nucleus and centrosomes in dividing cells. By combining mass spectrometry, biochemical and cell biological approaches, we show that the nuclear pore complex (NPC) component RanBP2 directly binds to BICD2 and recruits it to NPCs specifically in G2 phase of the cell cycle. BICD2, in turn, recruits dynein-dynactin to NPCs and as such is needed to keep centrosomes closely tethered to the nucleus prior to mitotic entry. When dynein function is suppressed by RNA interference-mediated depletion or antibody microinjection, centrosomes and nuclei are actively pushed apart in late G2 and we show that this is due to the action of kinesin-1. Surprisingly, depletion of BICD2 inhibits both dynein and kinesin-1-dependent movements of the nucleus and cytoplasmic NPCs, demonstrating that BICD2 is needed not only for the dynein function at the nuclear pores but also for the antagonistic activity of kinesin-1. Our study demonstrates that the nucleus is subject to opposing activities of dynein and kinesin-1 motors and that BICD2 contributes to nuclear and centrosomal positioning prior to mitotic entry through regulation of both dynein and kinesin-1.
authors.affiliation: Department of Cell Biology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Daniël
authors.initials: D
authors.lastname: Splinter
authors.forename: Marvin E
authors.initials: ME
authors.lastname: Tanenbaum
authors.forename: Arne
authors.initials: A
authors.lastname: Lindqvist
authors.forename: Dick
authors.initials: D
authors.lastname: Jaarsma
authors.forename: Annette
authors.initials: A
authors.lastname: Flotho
authors.forename: Ka Lou
authors.initials: KL
authors.lastname: Yu
authors.forename: Ilya
authors.initials: I
authors.lastname: Grigoriev
authors.forename: Dieuwke
authors.initials: D
authors.lastname: Engelsma
authors.forename: Elize D
authors.initials: ED
authors.lastname: Haasdijk
authors.forename: Nanda
authors.initials: N
authors.lastname: Keijzer
authors.forename: Jeroen
authors.initials: J
authors.lastname: Demmers
authors.forename: Maarten
authors.initials: M
authors.lastname: Fornerod
authors.forename: Frauke
authors.initials: F
authors.lastname: Melchior
authors.forename: Casper C
authors.initials: CC
authors.lastname: Hoogenraad
authors.forename: René H
authors.initials: RH
authors.lastname: Medema
authors.forename: Anna
authors.initials: A
authors.lastname: Akhmanova
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism* / Q000648:ultrastructure; D018385:Centrosome / Q000378:metabolism*; D000072159:Dynactin Complex; D004398:Dyneins / Q000378:metabolism*; D006801:Humans; D016547:Kinesins / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008938:Mitosis / Q000502:physiology*; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D022022:Nuclear Pore / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D008941:Spindle Apparatus / Q000378:metabolism; D020798:Two-Hybrid System Techniques
chemical_list: C435206:Bicd2 protein, mouse; D002352:Carrier Proteins; D000072159:Dynactin Complex; C470243:KIF11 protein, human; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D018832:Molecular Chaperones; D028861:Nuclear Pore Complex Proteins; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; C094493:ran-binding protein 2; D004398:Dyneins; D016547:Kinesins
references.citation: Curr Opin Cell Biol. 1999 Feb;11(1):45-53
references.pmid: 10047518
references.citation: Ann Anat. 2005 Nov;187(5-6):539-53
references.pmid: 16320833
references.citation: Curr Opin Cell Biol. 2002 Feb;14(1):44-9
references.pmid: 11792543
references.citation: J Biol Chem. 2001 Nov 9;276(45):41594-602
references.pmid: 11553612
references.citation: J Cell Biol. 2004 Sep 27;166(7):1003-14
references.pmid: 15381688
references.citation: J Cell Biol. 1996 Dec;135(6 Pt 1):1457-70
references.pmid: 8978815
references.citation: Nature. 2001 Dec 6;414(6864):611-6
references.pmid: 11740552
references.citation: EMBO Rep. 2009 May;10(5):480-6
references.pmid: 19305391
references.citation: Cell. 2003 Dec 26;115(7):825-36
references.pmid: 14697201
references.citation: Development. 2007 Oct;134(19):3419-25
references.pmid: 17827179
references.citation: Curr Biol. 2004 Apr 6;14(7):611-7
references.pmid: 15062103
references.citation: Cell. 2002 Jan 11;108(1):83-96
references.pmid: 11792323
references.citation: Cell. 2002 Jan 11;108(1):109-20
references.pmid: 11792325
references.citation: Cell Tissue Res. 1996 May;284(2):177-91
references.pmid: 8625385
references.citation: EMBO J. 2008 Dec 17;27(24):3235-45
references.pmid: 19020519
references.citation: Neuron. 2005 May 5;46(3):383-8
references.pmid: 15882636
references.citation: Traffic. 2005 Dec;6(12):1070-7
references.pmid: 16262719
references.citation: Cell. 1997 Jan 10;88(1):97-107
references.pmid: 9019411
references.citation: Int Rev Cytol. 1992;133:43-120
references.pmid: 1374369
references.citation: EMBO J. 2006 Jan 11;25(1):45-57
references.pmid: 16362039
references.citation: Genes Dev. 1997 Feb 15;11(4):423-35
references.pmid: 9042857
references.citation: Nat Cell Biol. 2007 Jul;9(7):788-96
references.pmid: 17546040
references.citation: Traffic. 2007 Dec;8(12):1722-1735
references.pmid: 17887960
references.citation: J Cell Biol. 2005 Dec 5;171(5):799-810
references.pmid: 16330710
references.citation: J Cell Biol. 2008 Sep 22;182(6):1063-71
references.pmid: 18809722
references.citation: J Cell Sci. 2004 Jan 15;117(Pt 2):133-41
references.pmid: 14676269
references.citation: Curr Biol. 2002 Oct 15;12(20):1792-5
references.pmid: 12401177
references.citation: PLoS One. 2008;3(11):e3763
references.pmid: 19018277
references.citation: EMBO J. 2001 Aug 1;20(15):4041-54
references.pmid: 11483508
references.citation: Curr Biol. 2006 Jul 25;16(14):1447-52
references.pmid: 16860745
references.citation: Trends Cell Biol. 2002 Jan;12(1):21-7
references.pmid: 11854006
references.citation: Mol Biol Cell. 2008 May;19(5):2300-10
references.pmid: 18305100
references.citation: Nat Cell Biol. 2005 Mar;7(3):219-22
references.pmid: 15738974
references.citation: J Cell Biol. 1999 Oct 18;147(2):321-34
references.pmid: 10525538
references.citation: Neuron. 2009 Oct 29;64(2):173-87
references.pmid: 19874786
references.citation: Transgenic Res. 2005 Aug;14(4):477-82
references.pmid: 16201414
references.citation: Mol Cell Biol. 2006 Sep;26(18):6772-85
references.pmid: 16943420
references.citation: Exp Cell Res. 2000 Aug 25;259(1):180-90
references.pmid: 10942590
references.citation: Nat Cell Biol. 2002 Dec;4(12):986-92
references.pmid: 12447383
references.citation: EMBO J. 2003 Nov 17;22(22):6004-15
references.pmid: 14609947
references.citation: Cell. 2000 Aug 4;102(3):279-91
references.pmid: 10975519
references.citation: J Cell Biol. 2007 Aug 27;178(5):785-98
references.pmid: 17724119
references.citation: Curr Biol. 1998 Apr 23;8(9):541-4
references.pmid: 9560347
references.citation: Genes Dev. 2009 Jul 1;23(13):1546-58
references.pmid: 19515976
references.citation: Annu Rev Cell Dev Biol. 1999;15:607-60
references.pmid: 10611974
references.citation: Nature. 1995 Jul 13;376(6536):184-8
references.pmid: 7603572
references.citation: Dev Cell. 2006 Jul;11(1):21-32
references.pmid: 16824950
references.citation: J Cell Sci. 2006 Dec 15;119(Pt 24):5021-9
references.pmid: 17158909
references.citation: Curr Biol. 2008 Feb 12;18(3):177-82
references.pmid: 18249114
references.citation: Dev Cell. 2007 Aug;13(2):305-14
references.pmid: 17681140
references.citation: J Cell Biol. 2005 Jan 3;168(1):141-53
references.pmid: 15631994
references.citation: Phys Biol. 2004 Jun;1(1-2):R1-11
references.pmid: 16204815
references.citation: Cell. 1986 Jun 6;45(5):699-709
references.pmid: 3518946
references.citation: Cell. 2002 Jan 11;108(1):97-107
references.pmid: 11792324
references.citation: J Cell Biol. 1999 Aug 9;146(3):597-608
references.pmid: 10444068
references.citation: J Cell Sci. 2007 Oct 1;120(Pt 19):3384-94
references.pmid: 17881500
references.citation: Nat Biotechnol. 2004 Dec;22(12):1567-72
references.pmid: 15558047
references.citation: J Biol Chem. 1995 Jun 9;270(23):14209-13
references.pmid: 7775481
references.citation: Nature. 2005 May 5;435(7038):114-8
references.pmid: 15875026
references.citation: Biochem Soc Trans. 2008 Dec;36(Pt 6):1368-72
references.pmid: 19021557
references.citation: Cell. 2003 Feb 21;112(4):467-80
references.pmid: 12600311
references.citation: J Cell Biol. 1999 Oct 4;147(1):135-50
references.pmid: 10508861
references.citation: Proc Natl Acad Sci U S A. 2003 Jun 24;100(13):7480-5
references.pmid: 12802011
references.citation: Curr Biol. 2004 Jul 13;14(13):R525-37
references.pmid: 15242636
references.citation: Proc Natl Acad Sci U S A. 2009 Feb 17;106(7):2194-9
references.pmid: 19164528
references.citation: Annu Rev Cell Dev Biol. 2005;21:347-80
references.pmid: 16212499
references.citation: Biochim Biophys Acta. 2008 Sep;1786(1):32-40
references.pmid: 18549824