ABA (phytohormone avscisic acid) : phtocormone growth inhibitor C 15 H 20 O 4 (M.W= 264.32)

1. Homeodomain protein ATHB6 is a target of the protein phosphatase ABI1 and regulates hormone responses in Arabidopsis

2. ABA (phytohormone avscisic acid) : phtocormone growth inhibitor C15H20O4 (M.W= 264.32) 1964, P.F Wareing isolated dormancy-inducible factor from Betula Pubescence ⇒ dormin

3. Function of ABA : role in regulating plant responses to adverse environmental cues – water shortage high osmolarity ⇒ elevation of plant low temperature ABA level leads to physiological adaptations ⇒ stomatal closure growth inhibition differential gene regulation for metabolic developmental adjustment

4. PP2C : protein serine/threonin phosphatase of PPM family present in both eukaryotes and prokaryotes no similarity seqencing structurally similar with PPP family consists of two domains ⇒ N-terminal catalytic domain - 6α-helices, 11β –sheets - two Mn+ coordnated by four aspartate, Glu residue ⇒ 90 residue C-terminal domainof 3α-helices

5. ABI1 : ABA-INSENSITIVE encodes a member of the 2C class of protein serine/threonine phosphatase negative regulator of abscisic acid signaling ATHB6 : isolated in low stringency screen for homeobox contain genes by use ATHB6 cDNA as a probe encodes a class 1 homeodomain-leucine zipper protein

6. Homepdomain-leucine zipper protein ⇓ approximately 60 amino acid residues producing either three or four α-helices and N-terminal arms : HD-Zip protein large family of transcription factors apparently unique to plant contain a leucine zipper domain C-terminal to the domain actives as protein dimers in the binding of DNA

7. Arabidopsis thaliana genome • Contanis 42 HD-Zip encoding gene • Designated Arabidopsis thaliana • homeobox gene⇒ ATHBz • four classes • HD-Zip Ⅰ~Ⅳ • ATHB6 • Induced by water deficit conditions • by ABA indicating a role • in the ABA mediated drought stress response

8. ABI1 : protein phoosphatase 2C key componen of signal transduction in arabidopsis refulates diverse responses to phytohormone abscisic acid (ABA) ATHB6 : analysing proteins capable of interacting with ABI1 identified homeodomain protein ATHB6 regulater of the ABA signal pathway

9. interaction between ATHB6 and ABI1 • Intact protein phosphatase domain • N-terminal domain of ATHB6 containing DNA-binding site • recognizes a cis-element present in ATHB6 promoter • core motif (CAATTATTA) • mediated ATHB6, ABA-dependent gene expression • in protoplasts • controll transgenic plants containing a luciferase gene • negative regulator of the ABA signal • to act downstream of ABI1

10. Fig. 1.Interaction of ATHB6 with ABI1 in the yeast two-hybrid system and in vitro.

11. Fig. 2. Dependence of ABI1 and ATHB6 interaction on the functional catalytic domain of PP2C.

12. Fig. 3. Specific binding of ATHB6 to a cis-element present in the ATHB6 promoter.

13. Fig. 4. Requirement for a functional cis-element and ATHB6 for ABA-mediated promoter activation in Arabidopsis protoplasts.

14. Fig. 5. ABA- and ABI1-dependent activation of the ATHB6 promoter.

15. Fig. 6. Analysis of ABA responses in Arabidopsis plants with ectopic expression of ATHB6.