Thylakoid lumen carbonic anhydrase (CAH3) mutation suppresses air-Dier phenotype of LCIB mutant in Chlamydomonas reinhardtii.
An active CO2-concentrating mechanism is induced when Chlamydomonas reinhardtii acclimates to limiting inorganic carbon (Ci), either low-CO2 (L-CO2; air level; approximately 0.04% CO2) or very low-CO2 (VL-CO2; approximately 0.01% CO2) conditions. A mutant, ad1, which is defective in the limiting-CO2-inducible, plastid-localized LCIB, can grow in high-CO2 or VL-CO2 conditions but dies in L-CO2, indicating a deficiency in a L-CO2-specific Ci uptake and accumulation system. In this study, we identified two ad1 suppressors that can grow in L-CO2 but die in VL-CO2. Molecular analyses revealed that both suppressors have mutations in the CAH3 gene, which encodes a thylakoid lumen localized carbonic anhydrase. Photosynthetic rates of L-CO2-acclimated suppressors under acclimation CO2 concentrations were more than 2-fold higher than ad1, apparently resulting from a more than 20-fold increase in the intracellular concentration of Ci as measured by direct Ci uptake. However, photosynthetic rates of VL-CO2-acclimated cells under acclimation CO2 concentrations were too low to support growth in spite of a significantly elevated intracellular Ci concentration. We conclude that LCIB functions downstream of CAH3 in the CO2-concentrating mechanism and probably plays a role in trapping CO2 released by CAH3 dehydration of accumulated Ci. Apparently dehydration by the chloroplast stromal carbonic anhydrase CAH6 of the very high internal Ci caused by the defect in CAH3 provides Rubisco sufficient CO2 to support growth in L-CO2-acclimated cells, but not in VL-CO2-acclimated cells, even in the absence of LCIB.
Duanmu
Deqiang
D
Department of Genetics, Development and Cell Biology, Iowa State University, Ames, Iowa 50011, USA.
Wang
Yingjun
Y
Spalding
Martin H
MH
D000818:Animals; D002244:Carbon / Q000378:metabolism; D002245:Carbon Dioxide / Q000378:metabolism; D002256:Carbonic Anhydrases / Q000235:genetics*; D016825:Chlamydomonas reinhardtii / Q000201:enzymology* / Q000235:genetics*; D003062:Codon / Q000235:genetics; D018388:Codon, Terminator / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D010101:Oxygen Consumption; D010641:Phenotype; D010788:Photosynthesis; D016133:Polymerase Chain Reaction; D013489:Suppression, Genetic; D020524:Thylakoids / Q000201:enzymology*
D003062:Codon; D018388:Codon, Terminator; D002245:Carbon Dioxide; D002244:Carbon; D002256:Carbonic Anhydrases