Identification of SUMO-2/3-modified proteins associated with mitotic chromosomes

Caelin Cubeñas-Potts, Tharan Srikumar, Christine Lee, Omoruyi Osula, Divya Subramonian, Xiang Dong Zhang, Robert J. Cotter, Brian Raught, Michael J. Matunis

Research output: Contribution to journalArticlepeer-review

Abstract

Sumoylation is essential for progression through mitosis, but the specific protein targets and functions remain poorly understood. In this study, we used chromosome spreads to more precisely define the localization of SUMO-2/3 (small ubiquitin-related modifier) to the inner centromere and protein scaffold of mitotic chromosomes. We also developed methods to immunopurify proteins modified by endogenous, untagged SUMO-2/3 from mitotic chromosomes. Using these methods, we identified 149 chromosome-associated SUMO-2/3 substrates by nLC-ESI-MS/MS. Approximately one-third of the identified proteins have reported functions in mitosis. Consistent with SUMO-2/3 immunolocalization, we identified known centromere- and kinetochore-associated proteins, as well as chromosome scaffold associated proteins. Notably, >30 proteins involved in chromatin modification or remodeling were identified. Our results provide insights into the roles of sumoylation as a regulator of chromatin structure and other diverse processes in mitosis. Furthermore, our purification and fractionation methodologies represent an important compliment to existing approaches to identify sumoylated proteins using exogenously expressed and tagged SUMOs.

Original languageEnglish (US)
Pages (from-to)763-772
Number of pages10
JournalProteomics
Volume15
Issue number4
DOIs
StatePublished - Feb 1 2015

Keywords

  • Cell biology
  • Chromosome
  • Mitosis
  • SUMO

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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