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  Alteration of GCN5 levels in maize reveals dynamic responses to manipulating histone acetylation

Bhat, R. A., Riehl, M., Santandrea, G., Velasco, R., Slocombe, S., Donn, G., et al. (2003). Alteration of GCN5 levels in maize reveals dynamic responses to manipulating histone acetylation. Plant Journal, 33(3), 455-469.

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 Creators:
Bhat, R. A.1, Author           
Riehl, M., Author
Santandrea, G., Author
Velasco, R.2, Author           
Slocombe, S.2, Author           
Donn, G., Author
Steinbiss, H. H.3, Author           
Thompson, R. D.2, Author           
Becker, H. A.2, Author           
Affiliations:
1Dept. of Plant Microbe Interactions (Paul Schulze-Lefert), MPI for Plant Breeding Research, Max Planck Society, ou_1113572              
2Dept. of Plant Breeding and Yield Physiology (Francesco Salamini), MPI for Plant Breeding Research, Max Planck Society, ou_1113570              
3Dept. of Genetic Principles of Plant Breeding (Jozef Schell), MPI for Plant Breeding Research, Max Planck Society, ou_1113567              

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Free keywords: histone acetyltransferase; co-activator; GCN5; ADA2; maize; Tricnostatin A
 Abstract: The role played by histone acetyltransferase (HAT), GCN5, in transcriptional co-activation has been analysed in detail in yeast and mammals. Here, we present the cloning and expression pattern of Zmgcn5 , the maize homologue. The enzymatic activity of the recombinant ZmGCN5 was analysed with histone and nucleosome substrates. In situ hybridisation of developing maize kernels using Zmgcn5 as probe shows that the transcript is concentrated in rapidly dividing cells. To investigate the role of ZmGCN5 in the transcription of specific plant genes, direct protein-protein interactions were tested. A cDNA clone encoding a putative interacting partner in GCN5-adapter complexes, ZmADA2, was isolated and the interaction between ZmGCN5 and ZmADA2 was confirmed by a GST-spin down experiment. Co-immunoprecipitation of the plant transcriptional activator Opaque-2 and ZmADA2 in nuclear extracts suggests ADA2/GCN5- containing complexes to mediate transcriptional activation by binding of this bZIP factor. For a more general analysis of the effects of histone acetylation on plant gene expression, 2500 ESTs spotted on filters were hybridised with cDNA probes derived either from maize cell lines treated with Trichostatin A (TSA), or from a transgenic line expressing the ZmGCN5 antisense transcript. Several sequences showing marked changes in abundance were confirmed by RNA blot analysis. Inhibition of histone deacetylation with TSA is accompanied by a decrease in the abundance of ZmGCN5 acetylase protein, but by increases in mRNAs for histones H2A, H2B, H3 and H4. The elevated histone mRNA levels were not reflected in increasing histone protein concentrations, suggesting hyperacetylated histones arising from TSA treatment may be preferentially degraded and substituted by de novo synthesised histones. The ZmGCN5 antisense material showed suppression of the endogenous ZmGCN5 transcript and the profiling analysis revealed increased mRNA levels for H2A, H2B and H4. Furthermore, in the antisense line, a reduction in the amount of the RPD3-type HD1B-I histone deacetylase protein was observed. A model for linked regulation of histone acetylation and histone mRNA transcription is discussed.

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Language(s): eng - English
 Dates: 2003-02
 Publication Status: Issued
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 28835
ISI: 000180851300004
 Degree: -

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Title: Plant Journal
  Alternative Title : Plant J.
Source Genre: Journal
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Publ. Info: -
Pages: - Volume / Issue: 33 (3) Sequence Number: - Start / End Page: 455 - 469 Identifier: ISSN: 0960-7412