A modified "cross-talk" between histone H2B Lys-120 ubiquitination and H3 Lys-79 methylation

Agus Darwanto, Matthew P. Curtis, Matthew Schrag, Wolff Kirsch, Peng Liu, Guoliang Xu, Jonathan W. Neidigh, Kangling Zhang

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Western blot analysis is currently the major method utilized for quantitatively assessing histone global modifications. However, there is a growing need to develop a highly specific, accurate, and multisite quantitative method. Herein, we report a liquid chromatography-tandem mass spectrometry-multiple reaction monitoring method to simultaneously quantify multisite modifications with unmatched specificity, sensitivity, and throughput. With one set of purification of histones by high pressure liquid chromatography or SDS-PAGE, nearly 20 modification sites including acetylation, propionylation, methylation, and ubiquitination were quantified within 2 h for two samples to be compared. Using this method, the relative levels of H2B ubiquitination and H3 Lys-79 methylation were quantified in the U937 human leukemia cell line, U937 derivative cell lines overexpressing anti-secretory factor 10 (AF10) and mutant AF10 with the deletion of the hDot1 binding domain OM-LZ. We found that H2B ubiquitination is inversely correlated with H3 Lys-79 methylation. Therefore, we propose that a catalytic and inhibitory loop mechanism may better describe the crosstalk relationship between H2B ubiquitination and H3 Lys-79 methylation.

Original languageEnglish (US)
Pages (from-to)21868-21876
Number of pages9
JournalJournal of Biological Chemistry
Volume285
Issue number28
DOIs
StatePublished - Jul 9 2010
Externally publishedYes

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Methylation
Ubiquitination
Histones
Histone Code
Cells
High pressure liquid chromatography
Cell Line
Acetylation
U937 Cells
Liquid chromatography
Crosstalk
Tandem Mass Spectrometry
Liquid Chromatography
Purification
Mass spectrometry
Polyacrylamide Gel Electrophoresis
Leukemia
Western Blotting
High Pressure Liquid Chromatography
Throughput

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

A modified "cross-talk" between histone H2B Lys-120 ubiquitination and H3 Lys-79 methylation. / Darwanto, Agus; Curtis, Matthew P.; Schrag, Matthew; Kirsch, Wolff; Liu, Peng; Xu, Guoliang; Neidigh, Jonathan W.; Zhang, Kangling.

In: Journal of Biological Chemistry, Vol. 285, No. 28, 09.07.2010, p. 21868-21876.

Research output: Contribution to journalArticle

Darwanto, A, Curtis, MP, Schrag, M, Kirsch, W, Liu, P, Xu, G, Neidigh, JW & Zhang, K 2010, 'A modified "cross-talk" between histone H2B Lys-120 ubiquitination and H3 Lys-79 methylation', Journal of Biological Chemistry, vol. 285, no. 28, pp. 21868-21876. https://doi.org/10.1074/jbc.M110.126813
Darwanto, Agus ; Curtis, Matthew P. ; Schrag, Matthew ; Kirsch, Wolff ; Liu, Peng ; Xu, Guoliang ; Neidigh, Jonathan W. ; Zhang, Kangling. / A modified "cross-talk" between histone H2B Lys-120 ubiquitination and H3 Lys-79 methylation. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 28. pp. 21868-21876.
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