Key phosphorylation events in Spc29 and Spc42 guide multiple steps of yeast centrosome duplication

Michele Haltiner Jones, Eileen T. O'Toole, Amy S. Fabritius, Eric G. Muller, Janet B. Meehl, Sue L. Jaspersen, Mark Winey

Research output: Contribution to journalArticle

Abstract

Phosphorylation modulates many cellular processes during cell cycle progression. The yeast centrosome (called the spindle pole body, SPB) is regulated by the protein kinases Mps1 and Cdc28/Cdk1 as it nucleates microtubules to separate chromosomes during mitosis. Previously we completed an SPB phosphoproteome, identifying 297 sites on 17 of the 18 SPB components. Here we describe mutagenic analysis of phosphorylation events on Spc29 and Spc42, two SPB core components that were shown in the phosphoproteome to be heavily phosphorylated. Mutagenesis at multiple sites in Spc29 and Spc42 suggests that much of the phosphorylation on these two proteins is not essential but enhances several steps of mitosis. Of the 65 sites examined on both proteins, phosphorylation of the Mps1 sites Spc29-T18 and Spc29-T240 was shown to be critical for function. Interestingly, these two sites primarily influence distinct successive steps; Spc29-T240 is important for the interaction of Spc29 with Spc42, likely during satellite formation, and Spc29-T18 facilitates insertion of the new SPB into the nuclear envelope and promotes anaphase spindle elongation. Phosphorylation sites within Cdk1 motifs affect function to varying degrees, but mutations only have significant effects in the presence of an MPS1 mutation, supporting a theme of coregulation by these two kinases.

Original languageEnglish (US)
Pages (from-to)2280-2291
Number of pages12
JournalMolecular Biology of the Cell
Volume29
Issue number19
DOIs
StatePublished - Sep 15 2018

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Spindle Pole Bodies
Centrosome
Yeasts
Phosphorylation
Mitosis
Anaphase
Mutation
Nuclear Envelope
Microtubules
Mutagenesis
Protein Kinases
Cell Cycle
Proteins
Phosphotransferases
Chromosomes

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Jones, M. H., O'Toole, E. T., Fabritius, A. S., Muller, E. G., Meehl, J. B., Jaspersen, S. L., & Winey, M. (2018). Key phosphorylation events in Spc29 and Spc42 guide multiple steps of yeast centrosome duplication. Molecular Biology of the Cell, 29(19), 2280-2291. https://doi.org/10.1091/mbc.E18-05-0296

Key phosphorylation events in Spc29 and Spc42 guide multiple steps of yeast centrosome duplication. / Jones, Michele Haltiner; O'Toole, Eileen T.; Fabritius, Amy S.; Muller, Eric G.; Meehl, Janet B.; Jaspersen, Sue L.; Winey, Mark.

In: Molecular Biology of the Cell, Vol. 29, No. 19, 15.09.2018, p. 2280-2291.

Research output: Contribution to journalArticle

Jones, MH, O'Toole, ET, Fabritius, AS, Muller, EG, Meehl, JB, Jaspersen, SL & Winey, M 2018, 'Key phosphorylation events in Spc29 and Spc42 guide multiple steps of yeast centrosome duplication', Molecular Biology of the Cell, vol. 29, no. 19, pp. 2280-2291. https://doi.org/10.1091/mbc.E18-05-0296
Jones MH, O'Toole ET, Fabritius AS, Muller EG, Meehl JB, Jaspersen SL et al. Key phosphorylation events in Spc29 and Spc42 guide multiple steps of yeast centrosome duplication. Molecular Biology of the Cell. 2018 Sep 15;29(19):2280-2291. https://doi.org/10.1091/mbc.E18-05-0296
Jones, Michele Haltiner ; O'Toole, Eileen T. ; Fabritius, Amy S. ; Muller, Eric G. ; Meehl, Janet B. ; Jaspersen, Sue L. ; Winey, Mark. / Key phosphorylation events in Spc29 and Spc42 guide multiple steps of yeast centrosome duplication. In: Molecular Biology of the Cell. 2018 ; Vol. 29, No. 19. pp. 2280-2291.
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