Genome-scale Arabidopsis promoter array identifies targets of the histone acetyltransferase GCN5.
We have assembled approximately 20 000 Arabidopsis thaliana promoter regions, compatible with functional studies that require cloning and with microarray applications. The promoter fragments can be captured as modular entry clones (MultiSite Gateway format) via site-specific recombinational cloning, and transferred into vectors of choice to investigate transcriptional networks. The fragments can also be amplified by PCR and printed on glass arrays. In combination with immunoprecipitation of protein-DNA complexes (ChIP-chip), these arrays enable characterization of binding sites for chromatin-associated proteins or the extent of chromatin modifications at genome scale. The Arabidopsis histone acetyltransferase GCN5 associated with 40% of the tested promoters. At most sites, binding did not depend on the integrity of the GCN5 bromodomain. However, the presence of the bromodomain was necessary for binding to 11% of the promoter regions, and correlated with acetylation of lysine 14 of histone H3 in these promoters. Combined analysis of ChIP-chip and transcriptomic data indicated that binding of GCN5 does not strictly correlate with gene activation. GCN5 has previously been shown to be required for light-regulated gene expression and growth, and we found that GCN5 targets were enriched in early light-responsive genes. Thus, in addition to its transcriptional activation function, GCN5 may play an important role in priming activation of inducible genes under non-induced conditions.
Benhamed
Moussa
M
Institut de Biotechnologie des Plantes, UMR 8618, Centre National de la Recherche Scientifique, Université de Paris Sud 11, 91405 Orsay, France.
Martin-Magniette
Marie-Laure
ML
Taconnat
Ludivine
L
Bitton
Frédérique
F
Servet
Caroline
C
De Clercq
Rebecca
R
De Meyer
Björn
B
Buysschaert
Caroline
C
Rombauts
Stéphane
S
Villarroel
Raimundo
R
Aubourg
Sébastien
S
Beynon
Jim
J
Bhalerao
Rishikesh P
RP
Coupland
George
G
Gruissem
Wilhelm
W
Menke
Frank L H
FL
Weisshaar
Bernd
B
Renou
Jean-Pierre
JP
Zhou
Dao-Xiu
DX
Hilson
Pierre
P
D000107:Acetylation; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D047369:Chromatin Immunoprecipitation; D018744:DNA, Plant / Q000235:genetics; D030561:Databases, Nucleic Acid; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D018745:Genome, Plant; D051548:Histone Acetyltransferases / Q000235:genetics* / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D008027:Light; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation; D000099224:p300-CBP-Associated Factor
D029681:Arabidopsis Proteins; D018744:DNA, Plant; D051548:Histone Acetyltransferases; D006657:Histones; D014157:Transcription Factors; C474636:GCN5 protein, Arabidopsis; D000099224:p300-CBP-Associated Factor