Phospho-regulation of kinetochore-microtubule attachments by the Aurora kinase Ipl1p.
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.
Cheeseman
Iain M
IM
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.
Anderson
Scott
S
Jwa
Miri
M
Green
Erin M
EM
Kang
Jung seog
Js
Yates
John R
JR
Chan
Clarence S M
CS
Drubin
David G
DG
Barnes
Georjana
G
D000595:Amino Acid Sequence; D064127:Aurora Kinases; D001665:Binding Sites; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000502:physiology; D020090:Chromosome Segregation; D016384:Consensus Sequence; D004252:DNA Mutational Analysis; D005656:Fungal Proteins / Q000502:physiology; D018386:Kinetochores / Q000502:physiology*; D013058:Mass Spectrometry; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000502:physiology*; D008938:Mitosis; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000502:physiology; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins; D004718:Saccharomycetales / Q000502:physiology
C066751:CHL1 protein, S cerevisiae; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; C116003:DAM1 protein, S cerevisiae; D005656:Fungal Proteins; D008869:Microtubule-Associated Proteins; C112472:NDC80 protein, S cerevisiae; D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; C491504:ARK1 protein, S cerevisiae; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases