The Arabidopsis SET-domain protein ASHR3 is involved in stamen development and interacts with the bHLH transcription factor ABORTED MICROSPORES (AMS)
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The Arabidopsis thaliana genome contains more than 30 genes encoding SET-domain proteins that are thought to be epigenetic regulators of gene expression and chromatin structure. SET-domain proteins can be divided into subgroups, and members of the Polycomb group (PcG) and trithorax group (trxG) have been shown to be important regulators of development. Both in animals and plants some of these proteins are components of multimeric protein complexes. Here, we have analyzed the Arabidopsis trxG protein ASHR3 which has a SET domain and pre- and post-SET domains similar to that of Ash1 in Drosophila. In addition to the SET domain, a divergent PHD finger is found in the N-terminus of the ASHR3 protein. As expected from SET-domain proteins involved in transcriptional activation, ASHR3 (coupled to GFP) localizes to euchromatin. A yeast two-hybrid screening revealed that the ASHR3 protein interacts with the putative basic helix-loop-helix (bHLH) transcription factor ABORTED MICROSPORES (AMS), which is involved in anther and stamen development in Arabidopsis. Deletion mapping indicated that both the PHD finger and the SET domain mediate the interaction between the two proteins. Overexpression of ASHR3 led in general to growth arrest, and specifically to degenerated anthers and male sterility. Expression analyses demonstrated that ASHR3 like AMS is expressed in the anther and in stamen filaments. We therefore propose that AMS can target ASHR3 to chromatin and regulate genes involved in stamen development and function.
KeywordsProtein-protein interaction Stamen development Over-expression phenotype
Yeast two hybrid
Green fluorescent protein
Thanks to Susanne Opravil in Thomas Jenuwein’s lab for providing recombinant nucleosomes. Many thanks to B. Sæther, R. Falleth, and Solveig H. Engebretsen for technical assistance. The Research Council of Norway has supported this work and V.A. (grant 146652/431), P.E.G. (grant 166057/V40) and I.S.M. (grant 129525/420). The work was facilitated by the services provided by the Norwegian Arabidopsis Research Centre (NARC, http://www.narc.no/), a part of the Research Council of Norway’s National Programme for Research in Functional Genomics (FUGE).
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