Efficient readout of posttranslational codes on the 50-residue tail of histone H3 by high-resolution MS/MS

Nertila Siuti, Neil L. Kelleher*

*Corresponding author for this work

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

Histone modifications are highly linked to DNA methylation and together they exert epigenetic control over many activities in the cell including gene transcription. Using a streamlined mass spectrometric approach to determine changes in modification states in the first 50 residues of histone H3, we found a decrease in the global methylation states of H3.1 at Lys 9, Lys 14, and Lys 27 after inhibition of DNA methyltransferases by 5-aza-2′-deoxycytidine. Collisional ion dissociation methods proved adequate to determine site-specific H3 posttranslational modifications (PTMs) because ample backbone bonds are cleaved between each modification site and PTMs were stable to MS/MS using threshold fragmentation in a linear ion trap (LTQ). Our assay allows for a quick profiling and site-specific interrogation of modification states on the first 50 residues of H3 and is directly applicable to H3.1, H3.2, or H3.3 using most OrbiTrap, FT ICR, or TOF mass spectrometers.

Original languageEnglish (US)
Pages (from-to)180-187
Number of pages8
JournalAnalytical Biochemistry
Volume396
Issue number2
DOIs
StatePublished - Jan 15 2010

Keywords

  • DNA methyltransferases
  • FTMS
  • Histone H3
  • Mass spectrometry

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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