Kinetochore microtubule dynamics and attachment stability are regulated by Hec1.
Mitotic cells face the challenging tasks of linking kinetochores to growing and shortening microtubules and actively regulating these dynamic attachments to produce accurate chromosome segregation. We report here that Ndc80/Hec1 functions in regulating kinetochore microtubule plus-end dynamics and attachment stability. Microinjection of an antibody to the N terminus of Hec1 suppresses both microtubule detachment and microtubule plus-end polymerization and depolymerization at kinetochores of PtK1 cells. Centromeres become hyperstretched, kinetochore fibers shorten from spindle poles, kinetochore microtubule attachment errors increase, and chromosomes severely mis-segregate. The N terminus of Hec1 is phosphorylated by Aurora B kinase in vitro, and cells expressing N-terminal nonphosphorylatable mutants of Hec1 exhibit an increase in merotelic attachments, hyperstretching of centromeres, and errors in chromosome segregation. These findings reveal a key role for the Hec1 N terminus in controlling dynamic behavior of kinetochore microtubules.
DeLuca
Jennifer G
JG
Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA. jgdeluca@email.unc.edu
Gall
Walter E
WE
Ciferri
Claudio
C
Cimini
Daniela
D
Musacchio
Andrea
A
Salmon
E D
ED
D000596:Amino Acids / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000276:immunology; D064107:Aurora Kinase B; D064127:Aurora Kinases; D001704:Biopolymers; D003598:Cytoskeletal Proteins; D018604:Epitope Mapping; D006367:HeLa Cells; D006801:Humans; D016547:Kinesins / Q000378:metabolism; D018386:Kinetochores / Q000378:metabolism*; D008677:Metaphase; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism*; D008954:Models, Biological; D009687:Nuclear Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D021381:Protein Transport; D008941:Spindle Apparatus / Q000378:metabolism
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D001704:Biopolymers; C497850:CLASP1 protein, human; D003598:Cytoskeletal Proteins; C091489:KIF2C protein, human; D008869:Microtubule-Associated Proteins; C120529:NDC80 protein, human; D009687:Nuclear Proteins; C484695:SPC24 protein, human; C576921:AURKB protein, human; D064107:Aurora Kinase B; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases; D016547:Kinesins