SPT16 ubiquitylation by DCAF14-CRL4 regulates FACT binding to histones

Cell Rep. 2022 Mar 22;38(12):110541. doi: 10.1016/j.celrep.2022.110541.

Abstract

The histone chaperone complex FACT comprises SPT16 and SSRP1 and contributes to DNA replication, transcription, and repair, but how it plays such various roles is unclear. Here, we show that human SPT16 is ubiquitylated at lysine-674 (K674) by the DCAF14-CRL4 ubiquitin ligase. K674 is located in the middle domain of SPT16, and the corresponding residue of the yeast ortholog is critical for binding to histone H3.1-H4. We show that the middle domain of human SPT16 binds to histone H3.1-H4 and that this binding is inhibited by K674 ubiquitylation. Cells with heterozygous knockin of a K674R mutant of SPT16 manifest reduction of both SPT16 ubiquitylation and H3.1 in chromatin, a reduced population in mid S phase, impaired proliferation, and increased susceptibility to S phase stress. Our data thus indicate that SPT16 ubiquitylation by DCAF14-CRL4 regulates FACT binding to histones and may thereby control DNA replication-coupled histone incorporation into chromatin.

Keywords: CP: Molecular Biology; CRL4; DCAF14; DNA replication; FACT; chromatin; histone chaperone; histone incorporation; ubiquitin ligase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin
  • DNA-Binding Proteins
  • High Mobility Group Proteins
  • Histone Chaperones
  • Histones*
  • Humans
  • Lysine
  • Receptors, Interleukin-17
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins*
  • Transcriptional Elongation Factors
  • Ubiquitin-Protein Ligases
  • Ubiquitination

Substances

  • Chromatin
  • DNA-Binding Proteins
  • FACT protein, S cerevisiae
  • High Mobility Group Proteins
  • Histone Chaperones
  • Histones
  • IL17RB protein, human
  • Receptors, Interleukin-17
  • SSRP1 protein, human
  • Saccharomyces cerevisiae Proteins
  • Transcriptional Elongation Factors
  • Ubiquitin-Protein Ligases
  • Lysine