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The Novel Myb Transcription Factor LCR1 Regulates the CO₂-Responsive Gene Cah1, Encoding a Periplasmic Carbonic Anhydrase in Chlamydomonas reinhardtii

Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Oiwake-cho, Sakyo-ku, Kyoto, 606-8502, Japan

¹ To whom correspondence should be addressed. E-mail fukuzawa{at}lif.kyoto-u.ac.jp; fax 81-75-753-6127.

Chlamydomonas reinhardtii acclimates to CO₂-limiting stress by inducing a set of genes for a carbon-concentrating mechanism (CCM). This set includes the gene Cah1, which encodes a periplasmic carbonic anhydrase. Although physiological aspects of CO₂ response have been extensively studied, regulatory components, such as transcription factors involved in the acclimation, have not been well described in eukaryotic microalgae. Using an arylsulfatase gene driven by the Cah1 promoter, a regulatory mutant of Cah1 was isolated and named lcr1 (for low-CO₂ stress response). The photosynthetic affinity for inorganic carbon of lcr1 was reduced compared with that of wild-type cells. Expression of three low-CO₂-inducible genes, Cah1, Lci1, and Lci6, were regulated by LCR1 as shown by cDNA array and RNA gel blot analyses. The Lcr1 gene encodes a protein of 602 amino acids containing a single Myb domain, which binds to the Cah1-promoter region. Expression of Lcr1 was induced by lowering CO₂ levels and controlled by the regulatory factor CCM1. These results suggest that LCR1 transmits the low CO₂ signal to at least three CO₂-responsive genes and then fully induces CCM.

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