HDAC6 Substrate Discovery Using Proteomics-Based Substrate Trapping: HDAC6 Deacetylates PRMT5 to Influence Methyltransferase Activity

ACS Chem Biol. 2021 Aug 20;16(8):1435-1444. doi: 10.1021/acschembio.1c00303. Epub 2021 Jul 27.

Abstract

Histone deacetylase 6 (HDAC6) is upregulated in a variety of tumor cell lines and has been linked to many cellular processes, such as cell signaling, protein degradation, cell survival, and cell motility. HDAC6 is an enzyme that deacetylates the acetyllysine residues of protein substrates, and the discovery of HDAC6 substrates, including tubulin, has revealed many roles of HDAC6 in cell biology. Unfortunately, among the wide variety of acetylated proteins in the cell, only a few are verified as HDAC6 substrates, which limits the full characterization of HDAC6 cellular functions. Substrate trapping mutants were recently established as a tool to discover unanticipated substrates of histone deacetylase 1 (HDAC1). In this study, we applied the trapping approach to identify potential HDAC6 substrates. Among the high confidence protein hits after trapping, protein arginine methyl transferase 5 (PRMT5) was successfully validated as a novel HDAC6 substrate. PRMT5 acetylation enhanced its methyltransferase activity and symmetrical dimethylation of downstream substrates, revealing possible crosstalk between acetylation and methylation. Substrate trapping represents a powerful, systematic, and unbiased approach to discover substrates of HDAC6.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylation
  • Catalytic Domain / genetics
  • DNA Helicases / metabolism
  • HEK293 Cells
  • Histone Deacetylase 6 / chemistry
  • Histone Deacetylase 6 / genetics
  • Histone Deacetylase 6 / metabolism*
  • Humans
  • Mutation
  • Poly-ADP-Ribose Binding Proteins / metabolism
  • Protein-Arginine N-Methyltransferases / chemistry
  • Protein-Arginine N-Methyltransferases / metabolism*
  • Proteomics / methods
  • RNA Helicases / metabolism
  • RNA Recognition Motif Proteins / metabolism

Substances

  • Poly-ADP-Ribose Binding Proteins
  • RNA Recognition Motif Proteins
  • PRMT5 protein, human
  • Protein-Arginine N-Methyltransferases
  • HDAC6 protein, human
  • Histone Deacetylase 6
  • DNA Helicases
  • G3BP1 protein, human
  • RNA Helicases