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PMID: 11827982 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The yeast protein kinase Mps1p is required for assembly of the integral spindle pole body component Spc42p.

The Journal of cell biology ·Vol. 156 ·No. 3 ·2002-02-04 ·Pages 453-65

Castillo AR, Meehl JB, Morgan G, Schutz-Geschwender A, Winey M

Abstract

Saccharomyces cerevisiae MPS1 encodes an essential protein kinase that has roles in spindle pole body (SPB) duplication and the spindle checkpoint. Previously characterized MPS1 mutants fail in both functions, leading to aberrant DNA segregation with lethal consequences. Here, we report the identification of a unique conditional allele, mps1-8, that is defective in SPB duplication but not the spindle checkpoint. The mutations in mps1-8 are in the noncatalytic region of MPS1, and analysis of the mutant protein indicates that Mps1-8p has wild-type kinase activity in vitro. A screen for dosage suppressors of the mps1-8 conditional growth phenotype identified the gene encoding the integral SPB component SPC42. Additional analysis revealed that mps1-8 exhibits synthetic growth defects when combined with certain mutant alleles of SPC42. An epitope-tagged version of Mps1p (Mps1p-myc) localizes to SPBs and kinetochores by immunofluorescence microscopy and immuno-EM analysis. This is consistent with the physical interaction we detect between Mps1p and Spc42p by coimmunoprecipitation. Spc42p is a substrate for Mps1p phosphorylation in vitro, and Spc42p phosphorylation is dependent on Mps1p in vivo. Finally, Spc42p assembly is abnormal in a mps1-1 mutant strain. We conclude that Mps1p regulates assembly of the integral SPB component Spc42p during SPB duplication.

MeSH Terms
Alleles Amino Acid Sequence/genetics Centrosome/enzymology,ultrastructure Cytoskeletal Proteins/genetics,metabolism Fluorescent Antibody Technique Gene Dosage Genes, cdc/physiology Kinetochores/enzymology,ultrastructure Microscopy, Electron Mitosis/genetics Mutation/physiology Phosphoproteins/genetics,metabolism Phosphorylation Protein Serine-Threonine Kinases/genetics,metabolism Protein Structure, Tertiary/genetics Protein-Tyrosine Kinases/genetics,metabolism Saccharomyces cerevisiae/enzymology,genetics,ultrastructure Saccharomyces cerevisiae Proteins Spindle Apparatus/enzymology,genetics,ultrastructure
Chemicals
Cytoskeletal Proteins Phosphoproteins SPC42 protein, S cerevisiae Saccharomyces cerevisiae Proteins Protein-Tyrosine Kinases Protein Serine-Threonine Kinases MPS1 protein, S cerevisiae
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Castillo Andrea R
MCD Biology, UCB 347, University of Colorado, Boulder CO 80309, USA.
Meehl Janet B
Morgan Garry
Schutz-Geschwender Amy
Winey Mark
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200111025
Published
2002-02-04
Epub
2002-00-04
Pages
453-65
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2173341
Subset
IM
Grants
NIGMS NIH HHS · F31 GM019566 · United States
NIGMS NIH HHS · R01 GM051312 · United States
NIGMS NIH HHS · GM19566 · United States
NIGMS NIH HHS · GM51312 · United States
External Links
PubMed source DOI: 10.1083/jcb.200111025 PMC Full Text View journal details
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