Assessing the Role of Paralog-Specific Sumoylation of HDAC1

Methods Mol Biol. 2017:1510:329-337. doi: 10.1007/978-1-4939-6527-4_24.

Abstract

Attachment of ubiquitin or ubiquitin-like (Ubl) modifiers, such as the small ubiquitin-related modifier SUMO, is a posttranslational modification (PTM) that reversibly regulates the function and the stability of target proteins. The SUMO paralogs SUMO1 and SUMO2/3, although sharing a common conjugation pathway, seem to play different roles in the cell. Many regulatory mechanisms, which contribute to SUMO-paralog-specific modification, have emerged. We have recently found that cell environment affects SUMO-paralog-specific sumoylation of HDAC1, whose conjugation to SUMO1 and not to SUMO2 facilitates its protein turnover. Here, we describe how to identify SUMO-paralog-specific conjugation of HDAC1 and how the different expression of SUMO E3 ligases in the cell plays an important role in this mechanism.

Keywords: HDAC; HDAC1; PIASy; PTM; SUMO1; SUMO2/3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western / methods
  • HeLa Cells
  • Histone Deacetylase 1 / genetics
  • Histone Deacetylase 1 / metabolism*
  • Humans
  • Immunoprecipitation / methods
  • Lysine / metabolism
  • MCF-7 Cells
  • Protein Processing, Post-Translational*
  • SUMO-1 Protein / genetics
  • SUMO-1 Protein / metabolism*
  • Small Ubiquitin-Related Modifier Proteins / genetics
  • Small Ubiquitin-Related Modifier Proteins / metabolism*
  • Sumoylation
  • Transfection
  • Ubiquitins / genetics
  • Ubiquitins / metabolism*

Substances

  • SUMO-1 Protein
  • SUMO1 protein, human
  • SUMO2 protein, human
  • SUMO3 protein, human
  • Small Ubiquitin-Related Modifier Proteins
  • Ubiquitins
  • HDAC1 protein, human
  • Histone Deacetylase 1
  • Lysine