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PMID: 15843429 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Functional analysis of human microtubule-based motor proteins, the kinesins and dyneins, in mitosis/cytokinesis using RNA interference.

Molecular biology of the cell ·Vol. 16 ·No. 7 ·2005-07-00 ·Pages 3187-99

Zhu C, Zhao J, Bibikova M, Leverson JD, Bossy-Wetzel E, Fan JB, Abraham RT, Jiang W

Abstract

Microtubule (MT)-based motor proteins, kinesins and dyneins, play important roles in multiple cellular processes including cell division. In this study, we describe the generation and use of an Escherichia coli RNase III-prepared human kinesin/dynein esiRNA library to systematically analyze the functions of all human kinesin/dynein MT motor proteins. Our results indicate that at least 12 kinesins are involved in mitosis and cytokinesis. Eg5 (a member of the kinesin-5 family), Kif2A (a member of the kinesin-13 family), and KifC1 (a member of the kinesin-14 family) are crucial for spindle formation; KifC1, MCAK (a member of the kinesin-13 family), CENP-E (a member of the kinesin-7 family), Kif14 (a member of the kinesin-3 family), Kif18 (a member of the kinesin-8 family), and Kid (a member of the kinesin-10 family) are required for chromosome congression and alignment; Kif4A and Kif4B (members of the kinesin-4 family) have roles in anaphase spindle dynamics; and Kif4A, Kif4B, MKLP1, and MKLP2 (members of the kinesin-6 family) are essential for cytokinesis. Using immunofluorescence analysis, time-lapse microscopy, and rescue experiments, we investigate the roles of these 12 kinesins in detail.

MeSH Terms
3' Untranslated Regions Anaphase Biochemistry/methods Chromosomal Proteins, Non-Histone/chemistry,metabolism Chromosomes/ultrastructure Cytokinesis Dyneins/chemistry Electrophoresis, Polyacrylamide Gel Escherichia coli/metabolism Gene Library Green Fluorescent Proteins/metabolism HeLa Cells Humans Kinesins/chemistry Kinetochores/ultrastructure Microscopy, Fluorescence Microtubules/chemistry,metabolism Mitosis Phenotype Plasmids/metabolism RNA Interference RNA, Small Interfering/metabolism Reverse Transcriptase Polymerase Chain Reaction Spindle Apparatus Time Factors Transfection
Chemicals
3' Untranslated Regions Chromosomal Proteins, Non-Histone RNA, Small Interfering Green Fluorescent Proteins Dyneins Kinesins
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Zhu Changjun
The Burnham Institute, La Jolla, CA 92037, USA.
Zhao Jian
Bibikova Marina
Leverson Joel D
Bossy-Wetzel Ella
Fan Jian-Bing
Abraham Robert T
Jiang Wei
References (70)
70 references, click to expand
  1. Mitosis: MCAK under the aura of Aurora B.
    Curr Biol. 2004 May 4;14(9):R346-8 PMID: 15120087
  2. Antitumor activity of a kinesin inhibitor.
    Cancer Res. 2004 May 1;64(9):3276-80 PMID: 15126370
  3. Many commonly used siRNAs risk off-target activity.
    Biochem Biophys Res Commun. 2004 Jun 18;319(1):256-63 PMID: 15158470
  4. Differentiation of cytoplasmic and meiotic spindle assembly MCAK functions by Aurora B-dependent phosphorylation.
    Mol Biol Cell. 2004 Jun;15(6):2895-906 PMID: 15064354
  5. The Kip3-like kinesin KipB moves along microtubules and determines spindle position during synchronized mitoses in Aspergillus nidulans hyphae.
    Eukaryot Cell. 2004 Jun;3(3):632-45 PMID: 15189985
  6. PRC1: a human mitotic spindle-associated CDK substrate protein required for cytokinesis.
    Mol Cell. 1998 Dec;2(6):877-85 PMID: 9885575
  7. Kinesins klp5(+) and klp6(+) are required for normal chromosome movement in mitosis.
    J Cell Sci. 2002 Mar 1;115(Pt 5):931-40 PMID: 11870212
  8. Searching for the middle ground: mechanisms of chromosome alignment during mitosis.
    J Cell Biol. 2002 May 13;157(4):551-6 PMID: 12011106
  9. Chromosome dynamics: new light on Aurora B kinase function.
    Curr Biol. 2002 Jul 9;12(13):R458-60 PMID: 12121637
  10. Short RNA duplexes produced by hydrolysis with Escherichia coli RNase III mediate effective RNA interference in mammalian cells.
    Proc Natl Acad Sci U S A. 2002 Jul 23;99(15):9942-7 PMID: 12096193
  11. Unstable kinetochore-microtubule capture and chromosomal instability following deletion of CENP-E.
    Dev Cell. 2002 Sep;3(3):351-65 PMID: 12361599
  12. Spindle-kinetochore attachment requires the combined action of Kin I-like Klp5/6 and Alp14/Dis1-MAPs in fission yeast.
    EMBO J. 2002 Nov 15;21(22):6015-24 PMID: 12426374
  13. Kinesin motors and disease.
    Trends Cell Biol. 2002 Dec;12(12):585-91 PMID: 12495847
  14. Centromeres and kinetochores: from epigenetics to mitotic checkpoint signaling.
    Cell. 2003 Feb 21;112(4):407-21 PMID: 12600307
  15. The molecular motor toolbox for intracellular transport.
    Cell. 2003 Feb 21;112(4):467-80 PMID: 12600311
  16. Expression profiling reveals off-target gene regulation by RNAi.
    Nat Biotechnol. 2003 Jun;21(6):635-7 PMID: 12754523
  17. Survivin is required for stable checkpoint activation in taxol-treated HeLa cells.
    J Cell Sci. 2003 Jul 15;116(Pt 14):2987-98 PMID: 12783991
  18. Activating and silencing the mitotic checkpoint through CENP-E-dependent activation/inactivation of BubR1.
    Cell. 2003 Jul 11;114(1):87-98 PMID: 12859900
  19. M phase phosphoprotein 1 is a human plus-end-directed kinesin-related protein required for cytokinesis.
    J Biol Chem. 2003 Jul 25;278(30):27844-52 PMID: 12740395
  20. Centromere-associated protein-E is essential for the mammalian mitotic checkpoint to prevent aneuploidy due to single chromosome loss.
    J Cell Biol. 2003 Aug 18;162(4):551-63 PMID: 12925705
  21. Phosphorylation of mitotic kinesin-like protein 2 by polo-like kinase 1 is required for cytokinesis.
    J Cell Biol. 2003 Sep 1;162(5):863-75 PMID: 12939256
  22. The roles of microtubule-based motor proteins in mitosis: comprehensive RNAi analysis in the Drosophila S2 cell line.
    J Cell Biol. 2003 Sep 15;162(6):1003-16 PMID: 12975346
  23. The Drosophila kinesin-like protein KLP67A is essential for mitotic and male meiotic spindle assembly.
    Mol Biol Cell. 2004 Jan;15(1):121-31 PMID: 13679514
  24. The chromokinesin, KLP3A, dives mitotic spindle pole separation during prometaphase and anaphase and facilitates chromatid motility.
    Mol Biol Cell. 2004 Jan;15(1):219-33 PMID: 14528012
  25. Aurora B regulates MCAK at the mitotic centromere.
    Dev Cell. 2004 Feb;6(2):253-68 PMID: 14960279
  26. Aurora B phosphorylates centromeric MCAK and regulates its localization and microtubule depolymerization activity.
    Curr Biol. 2004 Feb 17;14(4):273-86 PMID: 14972678
  27. Depletion of centromeric MCAK leads to chromosome congression and segregation defects due to improper kinetochore attachments.
    Mol Biol Cell. 2004 Mar;15(3):1146-59 PMID: 14699064
  28. Kinesin superfamily protein member 4 (KIF4) is localized to midzone and midbody in dividing cells.
    Exp Mol Med. 2004 Feb 29;36(1):93-7 PMID: 15031677
  29. Kin I kinesins are microtubule-destabilizing enzymes.
    Cell. 1999 Jan 8;96(1):69-78 PMID: 9989498
  30. Human BUBR1 is a mitotic checkpoint kinase that monitors CENP-E functions at kinetochores and binds the cyclosome/APC.
    J Cell Biol. 1999 Sep 6;146(5):941-54 PMID: 10477750
  31. The kinesin-related protein, HSET, opposes the activity of Eg5 and cross-links microtubules in the mammalian mitotic spindle.
    J Cell Biol. 1999 Oct 18;147(2):351-66 PMID: 10525540
  32. An endoribonuclease-prepared siRNA screen in human cells identifies genes essential for cell division.
    Nature. 2004 Dec 23;432(7020):1036-40 PMID: 15616564
  33. Cell cycle-dependent translocation of PRC1 on the spindle by Kif4 is essential for midzone formation and cytokinesis.
    Proc Natl Acad Sci U S A. 2005 Jan 11;102(2):343-8 PMID: 15625105
  34. Recruitment of MKLP1 to the spindle midzone/midbody by INCENP is essential for midbody formation and completion of cytokinesis in human cells.
    Biochem J. 2005 Jul 15;389(Pt 2):373-81 PMID: 15796717
  35. Role of the midbody matrix in cytokinesis: RNAi and genetic rescue analysis of the mammalian motor protein CHO1.
    Mol Biol Cell. 2004 Jul;15(7):3083-94 PMID: 15075367
  36. Drosophila Klp67A is required for proper chromosome congression and segregation during meiosis I.
    J Cell Sci. 2004 Jul 15;117(Pt 16):3669-77 PMID: 15252134
  37. Relocation of Aurora B from centromeres to the central spindle at the metaphase to anaphase transition requires MKlp2.
    J Cell Biol. 2004 Jul 19;166(2):167-72 PMID: 15263015
  38. Gene silencing of CENP-E by small interfering RNA in HeLa cells leads to missegregation of chromosomes after a mitotic delay.
    Mol Biol Cell. 2004 Aug;15(8):3771-81 PMID: 15181147
  39. The KinI kinesin Kif2a is required for bipolar spindle assembly through a functional relationship with MCAK.
    J Cell Biol. 2004 Aug 16;166(4):473-8 PMID: 15302853
  40. Dynein/dynactin regulate metaphase spindle length by targeting depolymerizing activities to spindle poles.
    J Cell Biol. 2004 Aug 16;166(4):465-71 PMID: 15314063
  41. Essential roles of KIF4 and its binding partner PRC1 in organized central spindle midzone formation.
    EMBO J. 2004 Aug 18;23(16):3237-48 PMID: 15297875
  42. Cell cycle regulation of central spindle assembly.
    Nature. 2004 Aug 19;430(7002):908-13 PMID: 15282614
  43. Human chromokinesin KIF4A functions in chromosome condensation and segregation.
    J Cell Biol. 2004 Aug 30;166(5):613-20 PMID: 15326200
  44. A standardized kinesin nomenclature.
    J Cell Biol. 2004 Oct 11;167(1):19-22 PMID: 15479732
  45. Phosphorylation by p34cdc2 regulates spindle association of human Eg5, a kinesin-related motor essential for bipolar spindle formation in vivo.
    Cell. 1995 Dec 29;83(7):1159-69 PMID: 8548803
  46. XKCM1: a Xenopus kinesin-related protein that regulates microtubule dynamics during mitotic spindle assembly.
    Cell. 1996 Jan 12;84(1):37-47 PMID: 8548824
  47. Molecular characterization of the 50-kD subunit of dynactin reveals function for the complex in chromosome alignment and spindle organization during mitosis.
    J Cell Biol. 1996 Feb;132(4):617-33 PMID: 8647893
  48. Oscillating mitotic newt lung cell kinetochores are, on average, under tension and rarely push.
    J Cell Sci. 1996 Dec;109 ( Pt 12):2823-31 PMID: 9013330
  49. Mutation of a gene for a Drosophila kinesin-like protein, Klp38B, leads to failure of cytokinesis.
    J Cell Sci. 1997 Apr;110 ( Pt 8):945-54 PMID: 9152020
  50. Mitotic spindle positioning in Saccharomyces cerevisiae is accomplished by antagonistically acting microtubule motor proteins.
    J Cell Biol. 1997 Sep 8;138(5):1041-53 PMID: 9281582
  51. A chromatin-associated kinesin-related protein required for normal mitotic chromosome segregation in Drosophila.
    J Cell Biol. 1997 Dec 15;139(6):1361-71 PMID: 9396743
  52. CENP-E function at kinetochores is essential for chromosome alignment.
    J Cell Biol. 1997 Dec 15;139(6):1373-82 PMID: 9396744
  53. Interaction of a Golgi-associated kinesin-like protein with Rab6.
    Science. 1998 Jan 23;279(5350):580-5 PMID: 9438855
  54. Mitotic centromere-associated kinesin is important for anaphase chromosome segregation.
    J Cell Biol. 1998 Aug 10;142(3):787-801 PMID: 9700166
  55. A model for the proposed roles of different microtubule-based motor proteins in establishing spindle bipolarity.
    Curr Biol. 1998 Jul 30-Aug 13;8(16):903-13 PMID: 9707401
  56. Characterization of the kinetochore binding domain of CENP-E reveals interactions with the kinetochore proteins CENP-F and hBUBR1.
    J Cell Biol. 1998 Oct 5;143(1):49-63 PMID: 9763420
  57. Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen.
    Science. 1999 Oct 29;286(5441):971-4 PMID: 10542155
  58. Mitotic motors in Saccharomyces cerevisiae.
    Biochim Biophys Acta. 2000 Mar 17;1496(1):99-116 PMID: 10722880
  59. The Xenopus chromokinesin Xkid is essential for metaphase chromosome alignment and must be degraded to allow anaphase chromosome movement.
    Cell. 2000 Aug 18;102(4):411-24 PMID: 10966104
  60. Xkid, a chromokinesin required for chromosome alignment on the metaphase plate.
    Cell. 2000 Aug 18;102(4):425-35 PMID: 10966105
  61. Probing spindle assembly mechanisms with monastrol, a small molecule inhibitor of the mitotic kinesin, Eg5.
    J Cell Biol. 2000 Sep 4;150(5):975-88 PMID: 10973989
  62. The Rab6-binding kinesin, Rab6-KIFL, is required for cytokinesis.
    EMBO J. 2000 Nov 1;19(21):5711-9 PMID: 11060022
  63. The human kinesin-like protein RB6K is under tight cell cycle control and is essential for cytokinesis.
    Mol Cell Biol. 2001 Apr;21(8):2944-55 PMID: 11283271
  64. All kinesin superfamily protein, KIF, genes in mouse and human.
    Proc Natl Acad Sci U S A. 2001 Jun 19;98(13):7004-11 PMID: 11416179
  65. CENP-E is essential for reliable bioriented spindle attachment, but chromosome alignment can be achieved via redundant mechanisms in mammalian cells.
    Mol Biol Cell. 2001 Sep;12(9):2776-89 PMID: 11553716
  66. Eg5 is static in bipolar spindles relative to tubulin: evidence for a static spindle matrix.
    J Cell Biol. 2001 Sep 17;154(6):1125-33 PMID: 11564753
  67. The chromokinesin Kid is necessary for chromosome arm orientation and oscillation, but not congression, on mitotic spindles.
    J Cell Biol. 2001 Sep 17;154(6):1135-46 PMID: 11564754
  68. Central spindle assembly and cytokinesis require a kinesin-like protein/RhoGAP complex with microtubule bundling activity.
    Dev Cell. 2002 Jan;2(1):41-54 PMID: 11782313
  69. Zebrafish mitotic kinesin-like protein 1 (Mklp1) functions in embryonic cytokinesis.
    Physiol Genomics. 2002 Feb 11;8(1):51-66 PMID: 11842131
  70. TOGp, the human homolog of XMAP215/Dis1, is required for centrosome integrity, spindle pole organization, and bipolar spindle assembly.
    Mol Biol Cell. 2004 Apr;15(4):1580-90 PMID: 14718566
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e05-02-0167
Published
2005-07-00
Epub
2005-00-20
Pages
3187-99
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1165403
Subset
IM
Grants
NIGMS NIH HHS · R01 GM067859 · United States
NIGMS NIH HHS · GM67859 · United States
External Links
PubMed source DOI: 10.1091/mbc.e05-02-0167 PMC Full Text View journal details
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