The chloroplast sulfate transport system in the green alga Chlamydomonas reinhardtii

The genome of the model unicellular green alga Chlamydomonas reinhardtii contains four distinct genes, SulP, SulP2, Sbp and Sabc, which together are postulated to encode a chloroplast envelope-localized sulfate transporter holocomplex. In this work, evidence is presented that regulation of expression of SulP2, Sbp and Sabc is specifically modulated by sulfur availability to the cells. Induction of transcription and higher steady-state levels of the respective mRNAs are reported under S-deprivation conditions. No such induction could be observed under N or P deprivation conditions. Expression, localization, and complex-association of the Sabc protein was specifically investigated using cellular and chloroplast fractionations, BN-PAGE, SDS-PAGE and Western blot analyses. It is shown that Sabc protein levels in the cells increased under S-deprivation conditions, consistent with the observed induction of Sabc gene transcription. It is further shown that the Sabc protein co-localizes with SulP to the chloroplast envelope. Blue-native PAGE followed by Western blot analysis revealed the presence of an apparent 380 kDa complex in C. reinhardtii, specifically recognized by polyclonal antibodies against SulP and Sabc. These results suggest the presence and function in C. reinhardtii of a Sbp-SulP-SulP2-Sabc chloroplast envelope sulfate transporter holocomplex.