In Vivo DNA Affinity Purification and Histone Deacetylase Inhibitor Treatment Proves the Role of Histone Acetylation in the Expression Regulation of High-Molecular-Weight Glutenin Genes

Csaba Éva, Kitti Szőke-Pázsi, Szabolcs Makai, Gyöngyvér Gell, Attila Fábián, Edina Poczkodi, Gábor Tóth, László Sági, László Tamás, Angéla Juhász

Research output: Contribution to journalArticle

Abstract

High-molecular-weight glutenin subunit (HMW GS) proteins are major components of the gluten matrix, which is the physical basis of bread-making in wheat. Epigenetic and transcriptional regulations of HMW GS genes were studied both in silico and in wet lab to understand their tissue (endosperm) specific expression. Our co-expressional network analysis identified key transcription factor (TF) genes that regulate HMW GS genes. We also show here that HMW GS genes are inhibited in vegetative tissues by histone deacetylation as revealed by strong GUS expression in vascular tissues of transgenic barley seedlings harbouring HMW GS gene promoter::uidA-reporter gene fusions upon treatment with a histone deacetylase inhibitor. A novel method termed in vivo DNA affinity purification (IP) has been developed here for the isolation of histones and transcription factors binding to target DNA regions. The technique is based on the biolistic introduction of biotinylated PCR probes amplified from HMW GS gene promoters into wheat leaves. Twenty-four hours later, the probe is cross-linked with interacting factors and subsequently re-purified from plant nuclear extracts. Many proteins, ribosomal proteins and histones have so far been isolated. No lysine-acetylated histone protein fragments were found which further highlight the inhibiting effect of histone deacetylation on HMW GS gene expression.

Original languageEnglish
Pages (from-to)750-763
Number of pages14
JournalPlant Molecular Biology Reporter
Volume36
Issue number5-6
DOIs
Publication statusPublished - Dec 1 2018

Keywords

  • Epigenetic regulation
  • Seed storage protein
  • Tissue specificity
  • Transcription factor

ASJC Scopus subject areas

  • Molecular Biology
  • Plant Science

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