title: Fission yeast kinesin-8 Klp5 and Klp6 are interdependent for mitotic nuclear retention and required for proper microtubule dynamics.
abstract: Fission yeast has two kinesin-8s, Klp5 and Klp6, which associate to form a heterocomplex. Here, we show that Klp5 and Klp6 are mutually dependent on each other for nuclear mitotic localization. During interphase, they are exported to the cytoplasm. In sharp contrast, during mitosis, Klp5 and Klp6 remain in the nucleus, which requires the existence of each counterpart. Canonical nuclear localization signal (NLS) is identified in the nonkinesin C-terminal regions. Intriguingly individual NLS mutants (NLSmut) exhibit loss-of-function phenotypes, suggesting that Klp5 and Klp6 enter the nucleus separately. Indeed, although neither Klp5-NLSmut nor Klp6-NLSmut enters the nucleus, wild-type Klp6 or Klp5, respectively, does so with different kinetics. In the absence of Klp5/6, microtubule catastrophe/rescue frequency and dynamicity are suppressed, whereas growth and shrinkage rates are least affected. Remarkably, chimera strains containing only the N-terminal Klp5 kinesin domains cannot disassemble interphase microtubules during mitosis, leading to the coexistence of cytoplasmic microtubules and nuclear spindles with massive chromosome missegregation. In this strain, a marked reduction of microtubule dynamism, even higher than in klp5/6 deletions, is evident. We propose that Klp5 and Klp6 play a vital role in promoting microtubule dynamics, which is essential for the spatiotemporal control of microtubule morphogenesis.
authors.affiliation: Laboratory of Cell Regulation Cancer Research UK, London Research Institute, Lincoln's Inn Fields Laboratories, London WC2A 3PX, United Kingdom.
authors.forename: Amy
authors.initials: A
authors.lastname: Unsworth
authors.forename: Hirohisa
authors.initials: H
authors.lastname: Masuda
authors.forename: Susheela
authors.initials: S
authors.lastname: Dhut
authors.forename: Takashi
authors.initials: T
authors.lastname: Toda
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000502:physiology; D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000378:metabolism*; D016547:Kinesins / Q000235:genetics / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D008938:Mitosis / Q000502:physiology*; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*; D008941:Spindle Apparatus / Q000378:metabolism
chemical_list: D008869:Microtubule-Associated Proteins; D019913:Nuclear Localization Signals; D011993:Recombinant Fusion Proteins; D029702:Schizosaccharomyces pombe Proteins; C469734:Klp5 protein, S pombe; C459102:Klp6 protein, S pombe; D016547:Kinesins
references.citation: Nat Cell Biol. 2006 Sep;8(9):957-62
references.pmid: 16906145
references.citation: J Biol Chem. 2004 Nov 19;279(47):49384-94
references.pmid: 15339927
references.citation: EMBO J. 2005 Aug 17;24(16):2931-43
references.pmid: 16079915
references.citation: Curr Biol. 2007 Mar 20;17(6):488-98
references.pmid: 17346968
references.citation: Nat Cell Biol. 2001 Jan;3(1):E17-21
references.pmid: 11146645
references.citation: Mol Biol Cell. 2006 Mar;17(3):1421-35
references.pmid: 16394105
references.citation: Annu Rev Cell Dev Biol. 1997;13:83-117
references.pmid: 9442869
references.citation: Biophys J. 2004 Sep;87(3):1795-804
references.pmid: 15345558
references.citation: Genetics. 2003 Oct;165(2):477-88
references.pmid: 14573463
references.citation: Biochem Biophys Res Commun. 2008 Apr 11;368(3):670-6
references.pmid: 18262494
references.citation: Cell Motil Cytoskeleton. 2008 Aug;65(8):626-40
references.pmid: 18553361
references.citation: Nature. 1999 Mar 18;398(6724):256-60
references.pmid: 10094050
references.citation: Yeast. 1998 Jul;14(10):943-51
references.pmid: 9717240
references.citation: Trends Cell Biol. 2008 Jul;18(7):307-10
references.pmid: 18513970
references.citation: Mol Cell. 2007 Apr 13;26(1):1-14
references.pmid: 17434122
references.citation: Methods Enzymol. 1991;194:795-823
references.pmid: 2005825
references.citation: Curr Opin Cell Biol. 2005 Feb;17(1):82-8
references.pmid: 15661523
references.citation: Curr Opin Cell Biol. 2007 Feb;19(1):31-5
references.pmid: 17184986
references.citation: Mol Biol Cell. 2005 Jul;16(7):3187-99
references.pmid: 15843429
references.citation: Curr Biol. 2004 Apr 6;14(7):548-59
references.pmid: 15062095
references.citation: Trends Cell Biol. 2005 Sep;15(9):467-76
references.pmid: 16084724
references.citation: Curr Biol. 2002 Apr 16;12(8):610-21
references.pmid: 11967147
references.citation: J Cell Sci. 2003 Dec 15;116(Pt 24):4891-903
references.pmid: 14625383
references.citation: J Cell Biol. 1995 Sep;130(6):1387-99
references.pmid: 7559760
references.citation: Mol Biol Cell. 2004 Apr;15(4):1609-22
references.pmid: 14742702
references.citation: Mol Biol Cell. 2005 Mar;16(3):1378-95
references.pmid: 15647375
references.citation: Yeast. 2005 May;22(7):583-91
references.pmid: 15942936
references.citation: EMBO J. 2001 Jul 2;20(13):3389-401
references.pmid: 11432827
references.citation: Dev Cell. 2008 Feb;14(2):252-62
references.pmid: 18267093
references.citation: J Cell Biol. 2004 Oct 11;167(1):19-22
references.pmid: 15479732
references.citation: Mol Biol Cell. 2001 Dec;12(12):3919-32
references.pmid: 11739790
references.citation: Mol Biol Cell. 2007 Aug;18(8):3094-104
references.pmid: 17553931
references.citation: Nature. 2007 May 17;447(7142):334-7
references.pmid: 17476213
references.citation: Genes Cells. 2000 Oct;5(10):777-87
references.pmid: 11029654
references.citation: EMBO J. 2002 Nov 15;21(22):6015-24
references.pmid: 12426374
references.citation: Annu Rev Cell Dev Biol. 1999;15:607-60
references.pmid: 10611974
references.citation: Curr Biol. 2000 Jun 29;10(13):766-75
references.pmid: 10898975
references.citation: J Cell Sci. 2002 Mar 1;115(Pt 5):931-40
references.pmid: 11870212
references.citation: Nature. 2005 May 19;435(7040):308-12
references.pmid: 15902249
references.citation: Curr Biol. 2007 Aug 21;17(16):R643-5
references.pmid: 17714654
references.citation: Annu Rev Cell Dev Biol. 2004;20:867-94
references.pmid: 15473863
references.citation: Cell. 1997 Sep 19;90(6):1051-60
references.pmid: 9323133
references.citation: Nature. 1997 Nov 20;390(6657):308-11
references.pmid: 9384386
references.citation: Trends Biochem Sci. 1991 Dec;16(12):478-81
references.pmid: 1664152
references.citation: Mol Cell. 2000 Jan;5(1):27-34
references.pmid: 10678166
references.citation: Cell. 2007 Mar 23;128(6):1161-72
references.pmid: 17382884
references.citation: Annu Rev Biochem. 1998;67:265-306
references.pmid: 9759490
references.citation: Nat Cell Biol. 2006 Sep;8(9):913-23
references.pmid: 16906148
references.citation: Nat Biotechnol. 2004 Dec;22(12):1567-72
references.pmid: 15558047
references.citation: Science. 1997 Oct 3;278(5335):141-4
references.pmid: 9311922
references.citation: Trends Cell Biol. 2005 Feb;15(2):112-20
references.pmid: 15695098
references.citation: Mol Biol Cell. 2005 Jun;16(6):2719-33
references.pmid: 15772152
references.citation: Nat Biotechnol. 2006 Jul;24(7):841-7
references.pmid: 16823372
references.citation: Trends Cell Biol. 2005 Mar;15(3):121-4
references.pmid: 15752974
references.citation: Genes Cells. 2006 Jul;11(7):815-27
references.pmid: 16824200
references.citation: Cell. 1985 Aug;42(1):39-50
references.pmid: 3926325
references.citation: EMBO J. 1999 Mar 15;18(6):1660-72
references.pmid: 10075936
references.citation: Traffic. 2005 Mar;6(3):173-86
references.pmid: 15702986
references.citation: Mol Cell. 2004 Aug 13;15(3):317-27
references.pmid: 15304213
references.citation: Genes Cells. 2006 Apr;11(4):337-52
references.pmid: 16611238
references.citation: Curr Opin Cell Biol. 2005 Feb;17(1):89-97
references.pmid: 15661524
references.citation: Biochemistry. 1993 Feb 9;32(5):1285-93
references.pmid: 8448138