Mitotic control of kinetochore-associated dynein and spindle orientation by human Spindly

J Cell Biol. 2009 Jun 1;185(5):859-74. doi: 10.1083/jcb.200812167. Epub 2009 May 25.

Abstract

Mitotic spindle formation and chromosome segregation depend critically on kinetochore-microtubule (KT-MT) interactions. A new protein, termed Spindly in Drosophila and SPDL-1 in C. elegans, was recently shown to regulate KT localization of dynein, but depletion phenotypes revealed striking differences, suggesting evolutionarily diverse roles of mitotic dynein. By characterizing the function of Spindly in human cells, we identify specific functions for KT dynein. We show that localization of human Spindly (hSpindly) to KTs is controlled by the Rod/Zw10/Zwilch (RZZ) complex and Aurora B. hSpindly depletion results in reduced inter-KT tension, unstable KT fibers, an extensive prometaphase delay, and severe chromosome misalignment. Moreover, depletion of hSpindly induces a striking spindle rotation, which can be rescued by co-depletion of dynein. However, in contrast to Drosophila, hSpindly depletion does not abolish the removal of MAD2 and ZW10 from KTs. Collectively, our data reveal hSpindly-mediated dynein functions and highlight a critical role of KT dynein in spindle orientation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aurora Kinase B
  • Aurora Kinases
  • Chromosomes, Human / metabolism
  • Chromosomes, Human / ultrastructure
  • Cytoskeletal Proteins
  • Dynactin Complex
  • Dyneins / metabolism*
  • HeLa Cells
  • Humans
  • Kinetochores / metabolism*
  • Microtubule-Associated Proteins / metabolism
  • Mitosis*
  • Nuclear Proteins / metabolism
  • Paclitaxel / pharmacology
  • Protein Serine-Threonine Kinases / physiology
  • Signal Transduction
  • Spindle Apparatus / metabolism*
  • Spindle Apparatus / ultrastructure

Substances

  • Cytoskeletal Proteins
  • Dynactin Complex
  • Microtubule-Associated Proteins
  • NDC80 protein, human
  • Nuclear Proteins
  • AURKB protein, human
  • Aurora Kinase B
  • Aurora Kinases
  • Protein Serine-Threonine Kinases
  • Dyneins
  • Paclitaxel