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

53 Scopus citations

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'A modified "cross-talk" between histone H2B Lys-120 ubiquitination and H3 Lys-79 methylation'. Together they form a unique fingerprint.

  • Cite this

    Darwanto, A., Curtis, M. P., Schrag, M., Kirsch, W., Liu, P., Xu, G., Neidigh, J. W., & Zhang, K. (2010). A modified "cross-talk" between histone H2B Lys-120 ubiquitination and H3 Lys-79 methylation. Journal of Biological Chemistry, 285(28), 21868-21876. https://doi.org/10.1074/jbc.M110.126813