A new approach combining LC-MS and multivariate statistical analysis for revealing changes in histone modification levels

被引:1
作者
Bilgraer, Raphael [1 ]
Gillet, Sylvie [1 ]
Gil, Sophie [2 ]
Evain-Brion, Daniele [2 ]
Laprevote, Olivier [1 ,3 ]
机构
[1] Univ Paris 05, Fac Sci Pharmaceut & Biol Paris, Sorbonne Paris Cite, UMR CNRS 8638, F-75006 Paris, France
[2] Univ Paris 05, Fac Sci Pharmaceut & Biol Paris, Sorbonne Paris Cite, INSERM U1139, F-75006 Paris, France
[3] Hop Lariboisiere, AP HP, Serv Toxicol Biol, F-75475 Paris 10, France
关键词
ELECTRON-CAPTURE DISSOCIATION; MASS-SPECTROMETRY; TOP-DOWN; PATTERN-RECOGNITION; MIDDLE-DOWN; CELL-LINE; DATA SETS; METABOLOMICS; ACETYLATION; PROTEOMICS;
D O I
10.1039/c4mb00395k
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
While acting upon chromatin compaction, histone post-translational modifications (PTMs) are involved in modulating gene expression through histone-DNA affinity and protein-protein interactions. These dynamic and environment-sensitive modifications are constitutive of the histone code that reflects the transient transcriptional state of the chromatin. Here we describe a global screening approach for revealing epigenetic disruption at the histone level. This original approach enables fast and reliable relative abundance comparison of histone PTMs and variants in human cells within a single LC-MS experiment. As a proof of concept, we exposed BeWo human choriocarcinoma cells to sodium butyrate (SB), a universal histone deacetylase (HDAC) inhibitor. Histone acid-extracts (n = 45) equally representing 3 distinct classes, Control, 1 mM and 2.5 mM SB, were analysed using ultra-performance liquid chromatography coupled with a hybrid quadrupole time-of-flight mass spectrometer (UPLC-QTOF-MS). Multivariate statistics allowed us to discriminate control from treated samples based on differences in their mass spectral profiles. Several acetylated and methylated forms of core histones emerged as markers of sodium butyrate treatment. Indeed, this untargeted histonomic approach could be a useful exploratory tool in many cases of xenobiotic exposure when histone code disruption is suspected.
引用
收藏
页码:2974 / 2983
页数:10
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