Structural and functional analysis of the C-terminal STAS domain of the Arabidopsis thaliana sulfate transporter SULTR1.2

H. Rouached (1) , P. Berthomieu (1) , E. El Kassis (1) , N. Cathala (1) , V. Catherinot (2) , G. Labesse (2) , J.C. Davidian (1) , P. Fourcroy (1)

1 BPMP - Biochimie et Physiologie Moléculaire des Plantes
2 CBS - Centre de Biochimie Structurale [Montpellier]

The C-terminal region of sulfate transporters from plants and animals belonging to the SLC26 family members shares a weak but significant similarity with the Bacillus sp. anti-anti-sigma protein SpoIIAA, thus defining the STAS domain (Sulfate Transporter and Anti Sigma antagonist). The present study is a structure/function analysis of the STAS domain of SULTR1.2, an Arabidopsis thaliana sulfate transporter. A 3-D model of the SULTR1.2 STAS domain was built, which indicated that it shares the SpoIIAA folds. Moreover, the phosphorylation site, which is necessary for SpoIIAA activity, is conserved in the SULTR1.2 STAS domain. The model was used to direct mutagenesis studies using a yeast mutant defective for sulfate transport. Truncation of the whole SULTR1.2 STAS domain resulted in the loss of sulfate transport function. Analyses of small deletions and mutations showed that the C-terminal tail of the SULTR1.2 STAS domain, and particularly two cysteine residues, plays an important role in sulfate transport by SULTR1.2. All the substitutions made at the putative phosphorylation site Thr587 led to a complete loss of the sulfate transport function of SULTR1.2. The reduction or suppression of sulfate transport of the SULTR1.2 mutants in yeast was not due to an incorrect targeting to the plasma membrane. Both our 3D modeling and mutational analyses strengthen the hypothesis that the SULTR1.2 STAS domain is involved in protein-protein interactions that could control sulfate transport.

Mots clés: Arabidopsis; SULTR1.2

Domaines: Biologie végétale

H. Rouached, P. Berthomieu, E. El Kassis, N. Cathala, V. Catherinot, et al.. Structural and functional analysis of the C-terminal STAS domain of the Arabidopsis thaliana sulfate transporter SULTR1.2. Journal of Biological Chemistry, 2005, 280 (16), pp.15976-15983. ⟨10.1074/jbc.M501635200⟩. ⟨hal-00086931⟩