The SUMO Protease SENP1 and the Chromatin Remodeler CHD3 Interact and Jointly Affect Chromatin Accessibility and Gene Expression

J Biol Chem. 2018 Oct 5;293(40):15439-15454. doi: 10.1074/jbc.RA118.002844. Epub 2018 Aug 6.

Abstract

The small ubiquitin-like modifier (SUMO) post-translationally modifies lysine residues of transcription factors and co-regulators and thereby contributes to an important layer of control of the activities of these transcriptional regulators. Likewise, deSUMOylation of these factors by the sentrin-specific proteases (SENPs) also plays a role in gene regulation, but whether SENPs functionally interact with other regulatory factors that control gene expression is unclear. In the present work, we focused on SENP1, specifically, on its role in activation of gene expression investigated through analysis of the SENP1 interactome, which revealed that SENP1 physically interacts with the chromatin remodeler chromodomain helicase DNA-binding protein 3 (CHD3). Using several additional methods, including GST pulldown and co-immunoprecipitation assays, we validated and mapped this interaction, and using CRISPR-Cas9-generated CHD3- and SENP1-KO cells (in the haploid HAP1 cell line), we investigated whether these two proteins are functionally linked in regulating chromatin remodeling and gene expression. Genome-wide ATAC-Seq analysis of the CHD3- and SENP1-KO cells revealed a large degree of overlap in differential chromatin openness between these two mutant cell lines. Moreover, motif analysis and comparison with ChIP-Seq profiles in K562 cells pointed to an association of CHD3 and SENP1 with CCCTC-binding factor (CTCF) and SUMOylated chromatin-associated factors. Lastly, genome-wide RNA-Seq also indicated that these two proteins co-regulate the expression of several genes. We propose that the functional link between chromatin remodeling by CHD3 and deSUMOylation by SENP1 uncovered here provides another level of control of gene expression.

Keywords: ATAC-seq; CHD3; HAP1; SENP1; SUMO protease; chromatin regulation; chromatin remodeling; small ubiquitin-like modifier (SUMO); sumoylation; transcription; transcription coregulator.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CCCTC-Binding Factor / genetics
  • CCCTC-Binding Factor / metabolism
  • COS Cells
  • CRISPR-Cas Systems
  • Cell Line, Tumor
  • Chlorocebus aethiops
  • Chromatin / chemistry*
  • Chromatin / metabolism
  • Chromatin / ultrastructure
  • Chromatin Assembly and Disassembly*
  • Cloning, Molecular
  • Cysteine Endopeptidases / genetics
  • Cysteine Endopeptidases / metabolism*
  • DNA Helicases / genetics
  • DNA Helicases / metabolism*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Editing / methods
  • Gene Expression
  • Genetic Vectors / chemistry
  • Genetic Vectors / metabolism
  • HEK293 Cells
  • Humans
  • K562 Cells
  • Leukocytes / metabolism
  • Leukocytes / pathology
  • Mi-2 Nucleosome Remodeling and Deacetylase Complex / genetics
  • Mi-2 Nucleosome Remodeling and Deacetylase Complex / metabolism*
  • Protein Binding
  • Protein Processing, Post-Translational*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • CCCTC-Binding Factor
  • CTCF protein, human
  • Chromatin
  • Recombinant Fusion Proteins
  • SENP1 protein, human
  • Cysteine Endopeptidases
  • Mi-2 Nucleosome Remodeling and Deacetylase Complex
  • DNA Helicases
  • CHD3 protein, human