Hydrogen peroxide induces Sp1 methylation and thereby suppresses cyclin B1 via recruitment of Suv39H1 and HDAC1 in cancer cells

Free Radic Biol Med. 2011 Dec 15;51(12):2309-18. doi: 10.1016/j.freeradbiomed.2011.10.001. Epub 2011 Oct 8.

Abstract

Sp1 is an important transcription factor for a number of genes that regulate cell growth and survival. Sp1 is an anchor protein that recruits other factors to regulate its target genes positively or negatively, but the mechanism of its functional switch by which positive or negative coregulators are recruited is not clear. In this study, we found that Sp1 could be methylated and that methylation was maintained by treatment with pargyline, a lysine-specific demethylase 1 (LSD1) inhibitor or knock LSD1 down directly. Hydrogen peroxide treatment increased the methylation of Sp1 and repressed Sp1 transcriptional activity. Investigation of the mechanism by which methylation decreased Sp1 activity found that methylation of Sp1 increased the recruitment of Su(var) 3-9 homologue 1(Suv39H1) and histone deacetylase 1 (HDAC1) to the cyclin B1 promoter, resulting in deacetylation and methylation of histone H3 and subsequent downregulation of cyclin B1. Finally, downregulation of cyclin B1 led to cell cycle arrest at the G2 phase. These results show that methylation of Sp1 causes it to act as a negative regulator by recruiting Suv39H1 and HDAC1 to induce chromatin remodeling. This finding that methylation acts as a functional switch provides new insight into the modulation of Sp1 transcriptional activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cyclin B1 / antagonists & inhibitors*
  • Cyclin B1 / genetics
  • Cyclin B1 / metabolism
  • HeLa Cells
  • Histone Deacetylase 1 / metabolism*
  • Humans
  • Hydrogen Peroxide / pharmacology*
  • Methylation
  • Methyltransferases / metabolism*
  • Repressor Proteins / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sp1 Transcription Factor / metabolism*
  • Tumor Cells, Cultured

Substances

  • Cyclin B1
  • Repressor Proteins
  • Sp1 Transcription Factor
  • Hydrogen Peroxide
  • SUV39H1 protein, human
  • Methyltransferases
  • HDAC1 protein, human
  • Histone Deacetylase 1