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Co-regulation proteomics reveals substrates and mechanisms of APC/C-dependent degradation

  • Sasha A. Singh
  • , Dominic Winter
  • , Marc Kirchner
  • , Ruchi Chauhan
  • , Saima Ahmed
  • , Nurhan Ozlu
  • , Amit Tzur
  • , Judith A. Steen
  • , Hanno Steen
  • Boston Children's Hospital
  • Brigham and Women’s Hospital
  • University of Bonn
  • Koc University

Research output: Contribution to journalArticlepeer-review

66 Scopus citations

Abstract

Using multiplexed quantitative proteomics, we analyzed cell cycledependent changes of the human proteome. We identified >4,400 proteins, each with a six-point abundance profile across the cell cycle. Hypothesizing that proteins with similar abundance profiles are co-regulated, we clustered the proteins with abundance profiles most similar to known Anaphase-Promoting Complex/Cyclosome (APC/C) substrates to identify additional putative APC/C substrates. This protein profile similarity screening (PPSS) analysis resulted in a shortlist enriched in kinases and kinesins. Biochemical studies on the kinesins confirmed KIFC1, KIF18A, KIF2C, and KIF4A as APC/C substrates. Furthermore, we showed that the APC/CCDH1- dependent degradation of KIFC1 regulates the bipolar spindle formation and proper cell division. A targeted quantitative proteomics experiment showed that KIFC1 degradation is modulated by a stabilizing CDK1-dependent phosphorylation site within the degradation motif of KIFC1. The regulation of KIFC1 (de-)phosphorylation and degradation provides insights into the fidelity and proper ordering of substrate degradation by the APC/C during mitosis.

Original languageEnglish
Pages (from-to)385-399
Number of pages15
JournalEMBO Journal
Volume33
Issue number4
DOIs
StatePublished - 18 Feb 2014

Funding

FundersFunder number
German Academic Exchange Service
National Institutes of HealthR01NS066973
National Institute of General Medical SciencesR01GM094844

    Keywords

    • Dynamic proteomics
    • Protein profile similarity screening
    • Quantitative proteomics
    • TMT-labeling
    • Ubiquitination-dependent protein degradation

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