Analyses of CIA5, the master regulator of the carbon-concentrating mechanism in Chlamydomonas reinhardtii, and its control of gene expression

In numerous studies, the CIA5 gene of Chlamydomonas reinhardtii Dangeard has been shown to control the expression of several CO2-responsive genes when cells are shifted to higher or lower levels of CO2. Using DNA microarray analyses with arrays containing 2764 unique cDNA sequences, we have demonstrated that several additional genes are controlled by the CIA5 gene, some increasing in expression when CO2 levels are lowered and others decreasing. Not all genes that respond to changes in CO2 concentrations are controlled by CIA5. For example, the RH1 gene, is markedly induced when both wild-type and cia5 mutant cells are shifted to high levels of CO2. We demonstrate that cycloheximide (an inhibitor of cytoplasmic protein synthesis) has no apparent effect on the initial induction of CO2-responsive genes, suggesting constitutive presence of all the molecular machinery needed by the cell to immediately respond to changes in CO2 levels. This observation is consistent with our earlier suggestions that CIA5 or another key component(s) of the carbon-concentrating mechanism must be rapidly activated (or inactivated) as part of the response of C. reinhardtii to changes in external CO2 levels. We present new, direct evidence that CIA5 is localized to the nucleus in both low- and high-CO2 conditions.