The Arabidopsis NPR1 protein is essential for regulating salicylic acid-dependent gene expression during systemic acquired resistance. NPR1 interacts differentially with members of the TGA class of basic domain/Leu zipper transcription factors and regulates their DNA binding activity. Here, we report that although TGA1 does not interact with NPR1 in yeast two-hybrid assays, treatment with salicylic acid induces the interaction between these proteins in Arabidopsis leaves. This phenomenon is correlated with a reduction of TGA1 Cys residues. Furthermore, site-directed mutagenesis of TGA1 Cys-260 and Cys-266 enables the interaction with NPR1 in yeast and Arabidopsis. Together, these results indicate that TGA1 relies on the oxidation state of Cys residues to mediate the interaction with NPR1. An intramolecular disulfide bridge in TGA1 precludes interaction with NPR1, and NPR1 can only stimulate the DNA binding activity of the reduced form of TGA1. Unlike its animal and yeast counterparts, the DNA binding activity of TGA1 is not redox regulated; however, this property is conferred by interaction with the NPR1 cofactor.
Abstract. We report the elk (Cervus elaphus) thyroid stimulating hormone (TSH) β-subunit cDNA cloning, nucleotide and deduced amino acid sequences. The TSH β-subunit cDNA was obtained by RT-PCR of polyadenylated pituitary RNA. The deduced elk TSH β-subunit peptide chain shares between 93 to 99% sequence similarities with the reported TSH β-subunit of a sub-set of related species. The TSH β-subunit gene is expressed in the elk pituitary gland as a mature transcript of approximately 600 bases, which corresponds to the size of the mRNA expressed in the sheep pituitary gland. Seasonal expression of the pituitary gonadotropin genes was investigated by Northern blot analyses. Samples of elk pituitary glands collected during the breeding season showed elevated steady state levels of common α-subunit and FSH and LH β-subunit gene expression, consistent with the seasonal reproductive cycling of this species. Samples collected before the breeding season demonstrated decreased expression of the gonadotropin genes. TSH, which is not directly tied to reproduction, had similar levels of expression, regardless of the animal's reproductive status. North American elk are temperate cervids and strict seasonal breeders, which start the mating season in autumn, in response to decreasing photoperiod [3]. Hence, most of their lifesustaining activities, such as feeding, locomotion, s l e e p a n d r e p r o d u c t i o n a r e p h o t o p e r i o d dependent. Activity of their reproductive organs, including testis, ovaries and accessory sex glands, exhibit substantial annual changes along with the circulatory levels of sex and pituitary hormones [4,5]. In the absence of pregnancy, deer hinds are polyestrous and are capable of exhibiting between four to nine continuous 17-19 day oestrous cycles over a 3-6 month period between autumn and spring [6][7][8]. Thus, the pituitary gland responding to photoperiod stimulus would change expression of the pituitary gonadotropin genes according to the reproductive season. We are interested in studying expression of the pituitary glycoprotein
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