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 language | English (US) |
|---|---|
| Pages (from-to) | 180-187 |
| Number of pages | 8 |
| Journal | Analytical Biochemistry |
| Volume | 396 |
| Issue number | 2 |
| DOIs | |
| State | Published - Jan 15 2010 |
Funding
The laboratory of N.L.K. was supported by the Packard Foundation , the Sloan Foundation , and the National Institutes of Health ( GM 067193-07 ).
Keywords
- DNA methyltransferases
- FTMS
- Histone H3
- Mass spectrometry
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology