Lysines Acetylome and Methylome Profiling of H3 and H4 Histones in Trichostatin A-Treated Stem Cells

Autor: Flora Cozzolino, Ilaria Iacobucci, Maria Chiara Monti, Vittoria Monaco, Tiziana Angrisano
Přispěvatelé: Cozzolino, F., Iacobucci, I., Monaco, V., Angrisano, T., Monti, M.
Rok vydání: 2021
Předmět:
Hydroxamic Acids
Hydroxamic Acid
lcsh:Chemistry
Histones
Mice
lcsh:QH301-705.5
TSA
Spectroscopy
mass spectrometry
biology
Chemistry
Peroxiredoxin
Acetylation
General Medicine
Methylation
Computer Science Applications
Chromatin
Cell biology
Histone
Peptide
DNA methylation
medicine.drug
Article
Catalysis
Inorganic Chemistry
Embryonic Stem Cell
medicine
Animals
Dimethyl Sulfoxide
Epigenetics
Amino Acid Sequence
Physical and Theoretical Chemistry
Molecular Biology
Embryonic Stem Cells
Animal
Lysine
Organic Chemistry
histone PTM
Peroxiredoxins
limited proteolysi
histone PTMs
Trichostatin A
lcsh:Biology (General)
lcsh:QD1-999
Gene Expression Regulation
Proteolysis
biology.protein
Histone deacetylase
activation of differentiation
Peptides
Octamer Transcription Factor-3
Protein Processing
Post-Translational
limited proteolysis
Zdroj: International Journal of Molecular Sciences
Volume 22
Issue 4
International Journal of Molecular Sciences, Vol 22, Iss 2063, p 2063 (2021)
ISSN: 1422-0067
Popis: Trichostatin A ([R-(E,E)]-7-[4-(dimethylamino) phenyl]-N-hydroxy- 4,6-dimethyl- 7-oxo-2,4-heptadienamide, TSA) affects chromatin state through its potent histone deacetylase inhibitory activity. Interfering with the removal of acetyl groups from lysine residues in histones is one of many epigenetic regulatory processes that control gene expression. Histone deacetylase inhibition drives cells toward the differentiation stage, favoring the activation of specific genes. In this paper, we investigated the effects of TSA on H3 and H4 lysine acetylome and methylome profiling in mice embryonic stem cells (ES14), treated with trichostatin A (TSA) by using a new, untargeted approach, consisting of trypsin-limited proteolysis experiments coupled with MALDI-MS and LC-MS/MS analyses. The method was firstly set up on standard chicken core histones to probe the optimized conditions in terms of enzyme:substrate (E:S) ratio and time of proteolysis and, then, applied to investigate the global variations of the acetylation and methylation state of lysine residues of H3 and H4 histone in the embryonic stem cells (ES14) stimulated by TSA and addressed to differentiation. The proposed strategy was found in its simplicity to be extremely effective in achieving the identification and relative quantification of some of the most significant epigenetic modifications, such as acetylation and lysine methylation. Therefore, we believe that it can be used with equal success in wider studies concerning the characterization of all epigenetic modifications.
Databáze: OpenAIRE
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