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, L. 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
JournalPlant Molecular Biology Reporter
DOIs
Publication statusAccepted/In press - Jan 1 2018

Fingerprint

histone deacetylase
Histone Deacetylase Inhibitors
glutenins
acetylation
Acetylation
histones
Histones
Molecular Weight
molecular weight
DNA
Genes
genes
Therapeutics
Triticum
Transcription Factors
transcription factors
Biolistics
promoter regions
biolistics
Endosperm

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Plant Science

Cite this

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

In: Plant Molecular Biology Reporter, 01.01.2018.

Research output: Contribution to journalArticle

Éva, Csaba ; Szőke-Pázsi, Kitti ; Makai, Szabolcs ; Gell, Gyöngyvér ; Fábián, Attila ; Poczkodi, Edina ; Tóth, Gábor ; Sági, László ; Tamás, L. ; Juhász, Angéla. / 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. In: Plant Molecular Biology Reporter. 2018.
@article{db2ff3d43d024d70a4f03dbf26145d01,
title = "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",
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.",
keywords = "Epigenetic regulation, Seed storage protein, Tissue specificity, Transcription factor",
author = "Csaba {\'E}va and Kitti Szőke-P{\'a}zsi and Szabolcs Makai and Gy{\"o}ngyv{\'e}r Gell and Attila F{\'a}bi{\'a}n and Edina Poczkodi and G{\'a}bor T{\'o}th and L{\'a}szl{\'o} S{\'a}gi and L. Tam{\'a}s and Ang{\'e}la Juh{\'a}sz",
year = "2018",
month = "1",
day = "1",
doi = "10.1007/s11105-018-1117-8",
language = "English",
journal = "Plant Molecular Biology Reporter",
issn = "0735-9640",
publisher = "Springer New York",

}

TY - JOUR

T1 - 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

AU - Éva, Csaba

AU - Szőke-Pázsi, Kitti

AU - Makai, Szabolcs

AU - Gell, Gyöngyvér

AU - Fábián, Attila

AU - Poczkodi, Edina

AU - Tóth, Gábor

AU - Sági, László

AU - Tamás, L.

AU - Juhász, Angéla

PY - 2018/1/1

Y1 - 2018/1/1

N2 - 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.

AB - 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.

KW - Epigenetic regulation

KW - Seed storage protein

KW - Tissue specificity

KW - Transcription factor

UR - http://www.scopus.com/inward/record.url?scp=85055572896&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85055572896&partnerID=8YFLogxK

U2 - 10.1007/s11105-018-1117-8

DO - 10.1007/s11105-018-1117-8

M3 - Article

JO - Plant Molecular Biology Reporter

JF - Plant Molecular Biology Reporter

SN - 0735-9640

ER -