First published online October 17, 2002; 10.1104/pp.009886
Plant Physiol, November 2002, Vol. 130, pp. 1516-1526
The as-1 Promoter Element Is an Oxidative
Stress-Responsive Element and Salicylic Acid Activates It via Oxidative
Species1
Virginia
Garretón,
Jorge
Carpinelli,
Xavier
Jordana, and
Loreto
Holuigue*
Departamento de Genética Molecular y Microbiología,
Facultad de Ciencias Biológicas, P. Universidad Católica de
Chile, Santiago, Chile
The activation sequence-1
(as-1)-like element found in the promoter of some
glutathione S-transferase (GST) genes,
has been previously described as a salicylic acid (SA)- and
auxin-responsive element. In this paper, we tested the hypothesis that
the activating effect of SA on the as-1 element is
mediated by oxidative species. Supporting this hypothesis, our results
show that the antioxidants dimethylthiourea (DMTU) and
3-t-butyl-4-hydroxy-anizole (BHA) inhibit the SA-induced transcription
of genes controlled by as-1 elements in tobacco
(Nicotiana tabacum) plants [i.e. GNT35
gene coding for a GST and
(as-1)4/ -glucuronidase
(GUS) reporter transgene]. DMTU and BHA also inhibit
SA-activated as-1-binding activity in nuclear extracts.
Further support for the hypothesis that the as-1 element
is activated by oxidative species comes from our result showing that
light potentiates the SA-induced activation of the as-1
element. Furthermore, methyl viologen, a known oxidative stress inducer
in plants, also activates the as-1 element. Increasing H2O2 levels by incubation with
H2O2 or with the catalase inhibitor 3-amino-1,2,5-triazole does not activate the
(as-1)4/GUS gene. On the
contrary, 3-amino-1,2,5-triazole inhibits the activating effect of SA
on the (as-1)4/GUS
gene. These results suggest that oxidative species other than
H2O2 mediate the activation of the as-1 element by SA. Our results also suggest that even
though the as-1 binding activity is stimulated by
oxidative species, this is not sufficient for the transactivation of
genes controlled by this element. The complex interplay between SA and
reactive oxygen species in the transcriptional activation of defense
genes is discussed.
1
This work was supported by Fondecyt-Conicyt,
Chile (grant nos. 8980005 and 2980065).
*
Corresponding author; e-mail lholuig{at}genes.bio.puc.cl; fax
56-2-222-5515.
© 2002 American Society of Plant Biologists
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