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Published on May 29, 2009; 10.1104/pp.109.135228

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Received January 13, 2009
Accepted May 18, 2009

The role of annexin 1 in drought stress in Arabidopsis thaliana

Dorota Konopka-Postupolska *, Greg Clark , Grazyna Goch , Janusz Debski , Krzysztof Floras , Araceli Cantero , Bartlomiej Fijolek , Stanley Roux , and Jacek Hennig

Institute of Biochemistry and Biophysics, Polish Academy of Sciences, 02-106 Warsaw, Poland; Molecular Cell & Developmental Biology, University of Texas, Austin, Texas 78712 USA

* Corresponding author; email: konopka{at}ibb.waw.pl.

Annexins act as targets of calcium signals in eukaryotic cells, and recent results suggest that they play an important role in plant stress responses. We found that in Arabidopsis thaliana AnnAt1 mRNA levels were up-regulated in leaves by most of the stress treatments applied. Plants overexpressing AnnAt1 protein were more drought-tolerant and knockout plants more drought-sensitive than Col-0 plants. We also observed that H2O2 accumulation in guard cells was reduced in overexpressing plants and increased in knockout plants both before and after treatment with ABA. Oxidative protection resulting from AnnAt1 overexpression could be due to the low level of intrinsic peroxidase activity exhibited by this protein in vitro, previously linked to a conserved histidine residue found in a peroxidase-like motif. However, analysis of a mutant AnnAt1-H40A protein in a bacterial complementation test and in peroxidase activity assays indicate that this residue is not critical to the ability of AnnAt1 to confer oxidative protection. To further examine the mechanism(s) linking AnnAt1 expression to stress resistance we analyzed the reactive S3 cluster to determine if it plays a role in AnnAt1 oligomerization and/or is the site for post-translational modification. We found that the two cysteine residues in this cluster do not form intra- or inter- molecular bonds but are highly susceptible to oxidation–driven S-glutathionylation which decreases the Ca2+ affinity of AnnAt1 in vitro. Moreover, S-glutathionylation of AnnAt1 occurs in planta after ABA treatment, which suggests that this modification could be important in regulating the cellular function of AnnAt1 during stress responses.




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