| Description: | Acetylation and deacetylation of histones are known to play important roles in regulating gene expression in eukaryotes. Histone acetyltransferases and histone deacetylases (HDACs) have been identified in large multisubunit complexes that target these enzymes to specific sites in nuclear DNA. In an attempt to identify auxin response mutants in Arabidopsis, we used natural and synthetic auxin response elements, linked to both the GUS reporter and a hygromycin resistance gene (HPH), in a genetic screen for plants with elevated expression of these transgenes. Five mutant lines with elevated expression of both the GUS and HPH transgenes were allelic, and the mutant gene was identified by fine mapping followed by sequence analysis. The gene encodes a HDAC, related to yeast RPD3, which we refer to as AtHDAC1. The purpose of this proposal is to identify genes that are regulated by AtHDAC1, using DNA microarrays. One of the mutant AtHDAC1 alleles contains a G to A transition in the second intron acceptor site from the 5' end of the gene. RNA gel blot analysis indicated that AtHDAC1 mRNA transcript levels are reduced in this mutant, and may be entirely absent. This mutant line has the strongest visible phenotype, characterized by increased branching and reduced fertility. Duplicate poly A+ RNA samples will be isolated from seedlings of this mutant line, and from wild-type Columbia, the ecotype from which the mutant was derived. A comparison of the microarray hybridization patterns between mutant and wild-type seedlings should indicate which genes other than the auxin regulated transgenes show altered expression in the mutant line. There have been few studies on the functions of HDACs and other chromatin remodelling enzymes in plants, and the identification of AtHDAC1 mutants in Arabidpsis provides us with an exciting opportunity to examine whether this enzyme has global or selective effects on gene expression. |
NASCArrays Information at The BAR
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