Functional analysis of HD2 histone deacetylase homologues in Arabidopsis thaliana

Post-translational modification of histones, in particular acetylation, is an important mechanism in the regulation of eukaryotic gene expression. His tone deacetylases are enzymes that remove acetyl groups from the core histo nes and play a key role in the repression of transcription. HD2 is a maize histone deacetylase, which shows no sequence homology to the histone deacet ylases identified from other eukaryotes. We have identified two putative HD P-like histone deacetylase cDNA clones, AtHD2A and AtHD2B, from Arabidopsis thaliana by screening the expressed sequence tag database. AtHD2A and AtHD 2B encode putative proteins of 246 and 305 amino acids, and share 44% and 4 6% amino acid identity to the maize HD2, respectively. Northern blot analys is indicated that AtHD2A was highly expressed in flowers and young siliques of Arabidopsis plants, whereas AtHD2B was widely expressed in stems, leave s, flowers and young siliques. AtHD2A repressed transcription when directed to a promoter containing GAL4 binding sites as a GAL4 fusion protein. Dele tion of the extended acidic domain or the domain containing predicted catal ytic residues of AtHD2A resulted in the loss of gene repression activity, r evealing the importance of both domains to AtHD2A function. Arabidopsis pla nts were transformed with a gene construct comprising an AtHD2A cDNA in the antisense orientation driven by a strong constitutive promoter, -394tCUP. Silencing of AtHD2A expression resulted in aborted seed development in tran sgenic Arabidopsis plants, suggesting that the AtHD2A gene product was impo rtant in the reproductive development of Arabidopsis thaliana.