N-formylation of lysine in histone proteins as a secondary modification arising from oxidative DNA damage

Proc Natl Acad Sci U S A. 2007 Jan 2;104(1):60-5. doi: 10.1073/pnas.0606775103. Epub 2006 Dec 26.


The posttranslational modification of histone and other chromatin proteins has a well recognized but poorly defined role in the physiology of gene expression. With implications for interfering with these epigenetic mechanisms, we now report the existence of a relatively abundant secondary modification of chromatin proteins, the N(6)-formylation of lysine that appears to be uniquely associated with histone and other nuclear proteins. Using both radiolabeling and sensitive bioanalytical methods, we demonstrate that the formyl moiety of 3'-formylphosphate residues arising from 5'-oxidation of deoxyribose in DNA, caused by the enediyne neocarzinostatin, for example, acylate the N(6)-amino groups of lysine side chains. A liquid chromatography (LC)-tandem mass spectrometry (MS) method was developed to quantify the resulting N(6)-formyl-lysine residues, which were observed to be present in unperturbed cells and all sources of histone proteins to the extent of 0.04-0.1% of all lysines in acid-soluble chromatin proteins including histones. Cells treated with neocarzinostatin showed a clear dose-response relationship for the formation of N(6)-formyl-lysine, with this nucleosome linker-selective DNA-cleaving agent causing selective N(6)-formylation of the linker histone H1. The N(6)-formyl-lysine residue appears to represent an endogenous histone secondary modification, one that bears chemical similarity to lysine N(6)-acetylation recognized as an important determinant of gene expression in mammalian cells. The N(6)-formyl modification of lysine may interfere with the signaling functions of lysine acetylation and methylation and thus contribute to the pathophysiology of oxidative and nitrosative stress.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Acetylation
  • Cells, Cultured
  • Chromatography, Liquid
  • DNA Damage*
  • Deoxyribose / metabolism
  • Histones / metabolism*
  • Humans
  • Lysine / metabolism*
  • Oxidation-Reduction
  • Tandem Mass Spectrometry


  • Histones
  • Deoxyribose
  • Lysine