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The green alga Chlamydomonas reinhardtii possesses a CO 2 concentrating mechanism (CCM) that helps in successful acclimation to low CO 2 conditions. Current models of the CCM postulate that a series of ion transporters bring HCO 3  - from outside the cell to the thylakoid lumen, where the carbonic anhydrase 3 (CAH3) dehydrates accumulated HCO 3  - to CO 2 , raising the CO 2 concentration for Ribulose bisphosphate carboxylase/oxygenase (Rubisco). Previously, HCO 3  - transporters have been identified at both the plasma membrane and the chloroplast envelope, but the transporter thought to be on the thylakoid membrane has not been identified. Three paralogous genes ( BST1 , BST2 , and BST3 ) belonging to the bestrophin family have been found to be up-regulated in low CO 2 conditions, and their expression is controlled by CIA5, a transcription factor that controls many CCM genes. YFP fusions demonstrate that all 3 proteins are located on the thylakoid membrane, and interactome studies indicate that they might associate with chloroplast CCM components. A single mutant defective in BST3 has near-normal growth on low CO 2 , indicating that the 3 bestrophin-like proteins may have redundant functions. Therefore, an RNA interference (RNAi) approach was adopted to reduce the expression of all 3 genes at once. RNAi mutants with reduced expression of BST1 - 3 were unable to grow at low CO 2 concentrations, exhibited a reduced affinity to inorganic carbon (C i ) compared with the wild-type cells, and showed reduced C i uptake. We propose that these bestrophin-like proteins are essential components of the CCM that deliver HCO 3  - accumulated in the chloroplast stroma to CAH3 inside the thylakoid lumen.

Thylakoid localized bestrophin-like proteins are essential for the CO 2 concentrating mechanism of Chlamydomonas reinhardtii

Thylakoid localized bestrophin-like proteins are essential for the CO ₂ concentrating mechanism of Chlamydomonas reinhardtii