A proteomic study of mitotic phase-Specific interactors of EB1 reveals a role for SXIP-Mediated protein interactions in anaphase onset

Naoka Tamura, Judith E. Simon, Arnab Nayak, Rajesh Shenoy, Noriko Hiroi, Viviane Boilot, Akira Funahashi, Viji M. Draviam

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Microtubules execute diverse mitotic events that are spatially and temporally separated; the underlying regulation is poorly understood. By combining drug treatments, large-scale immunoprecipitation and mass spectrometry, we report the first comprehensive map of mitotic phase-specific protein interactions of the microtubule-end binding protein, EB1. EB1 interacts with some, but not all, of its partners throughout mitosis. We show that the interaction of EB1 with Astrin-SKAP complex, a key regulator of chromosome segregation, is enhanced during prometaphase, compared to anaphase. We find that EB1 and EB3, another EB family member, can interact directly with SKAP, in an SXIP-motif dependent manner. Using an SXIP defective mutant that cannot interact with EB, we uncover two distinct pools of SKAP at spindle microtubules and kinetochores. We demonstrate the importance of SKAP's SXIP-motif in controlling microtubule growth rates and anaphase onset, without grossly disrupting spindle function. Thus, we provide the first comprehensive map of temporal changes in EB1 interactors during mitosis and highlight the importance of EB protein interactions in ensuring normal mitosis.

Original languageEnglish
Pages (from-to)155-169
Number of pages15
JournalBiology Open
Volume4
Issue number2
DOIs
Publication statusPublished - 2015 Feb 15

Fingerprint

Anaphase
anaphase
Mitosis
Microtubules
Proteomics
proteomics
microtubules
mitosis
Drug therapy
Chromosomes
Prometaphase
Mass spectrometry
Kinetochores
Chromosome Segregation
Carrier Proteins
Proteins
proteins
Immunoprecipitation
kinetochores
chromosome segregation

Keywords

  • Cell cortex
  • Kinetochore
  • Microtubule
  • Mitosis
  • Plus-end

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

A proteomic study of mitotic phase-Specific interactors of EB1 reveals a role for SXIP-Mediated protein interactions in anaphase onset. / Tamura, Naoka; Simon, Judith E.; Nayak, Arnab; Shenoy, Rajesh; Hiroi, Noriko; Boilot, Viviane; Funahashi, Akira; Draviam, Viji M.

In: Biology Open, Vol. 4, No. 2, 15.02.2015, p. 155-169.

Research output: Contribution to journalArticle

Tamura, Naoka ; Simon, Judith E. ; Nayak, Arnab ; Shenoy, Rajesh ; Hiroi, Noriko ; Boilot, Viviane ; Funahashi, Akira ; Draviam, Viji M. / A proteomic study of mitotic phase-Specific interactors of EB1 reveals a role for SXIP-Mediated protein interactions in anaphase onset. In: Biology Open. 2015 ; Vol. 4, No. 2. pp. 155-169.
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