Phospho-regulation of Kinetochore-Microtubule Attachments by the Aurora Kinase Ipl1p

Cell. 2002 Oct 18;111(2):163-72. doi: 10.1016/s0092-8674(02)00973-x.

Abstract

The Aurora kinase Ipl1p plays a crucial role in regulating kinetochore-microtubule attachments in budding yeast, but the underlying basis for this regulation is not known. To identify Ipl1p targets, we first purified 28 kinetochore proteins from yeast protein extracts. These studies identified five previously uncharacterized kinetochore proteins and defined two additional kinetochore subcomplexes. We then used mass spectrometry to identify 18 phosphorylation sites in 7 of these 28 proteins. Ten of these phosphorylation sites are targeted directly by Ipl1p, allowing us to identify a consensus phosphorylation site for an Aurora kinase. Our systematic mutational analysis of the Ipl1p phosphorylation sites demonstrated that the essential microtubule binding protein Dam1p is a key Ipl1p target for regulating kinetochore-microtubule attachments in vivo.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Aurora Kinases
  • Binding Sites
  • Cell Cycle Proteins / metabolism*
  • Chromosomal Proteins, Non-Histone / physiology
  • Chromosome Segregation
  • Consensus Sequence
  • DNA Mutational Analysis
  • Fungal Proteins / physiology
  • Kinetochores / physiology*
  • Mass Spectrometry
  • Microtubule-Associated Proteins / metabolism*
  • Microtubules / physiology*
  • Mitosis
  • Molecular Sequence Data
  • Nuclear Proteins / physiology
  • Phosphorylation*
  • Protein-Serine-Threonine Kinases / physiology*
  • Saccharomyces cerevisiae Proteins*
  • Saccharomycetales / physiology

Substances

  • CHL1 protein, S cerevisiae
  • Cell Cycle Proteins
  • Chromosomal Proteins, Non-Histone
  • DAM1 protein, S cerevisiae
  • Fungal Proteins
  • Microtubule-Associated Proteins
  • NDC80 protein, S cerevisiae
  • Nuclear Proteins
  • Saccharomyces cerevisiae Proteins
  • ARK1 protein, S cerevisiae
  • Aurora Kinases
  • Protein-Serine-Threonine Kinases