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Plant Physiol, December 1999, Vol. 121, pp. 1329-1337
CO2-Responsive Transcriptional Regulation of
CAH1 Encoding Carbonic Anhydrase Is Mediated by
Enhancer and Silencer Regions in Chlamydomonas
reinhardtii1
Ken-ichi
Kucho,
Kanji
Ohyama, and
Hideya
Fukuzawa*
Laboratory of Plant Molecular Biology, Division of Integrated Life
Science, Graduate School of Biostudies, Kyoto University, Kyoto
606-8502, Japan
Chlamydomonas
reinhardtii adapts to the stress of CO2-limiting
conditions through the induction of a set of genes including CAH1, which encodes a periplasmic carbonic anhydrase.
CAH1 is up-regulated under low-CO2
conditions (air containing 0.04% [v/v] CO2) in the
presence of light, whereas it is down-regulated under high-CO2 conditions (5% [v/v] CO2) or in the
dark. In an effort to identify cis-elements involved in the
transcriptional regulation of CAH1, a series of
5'-nested deletions of the region upstream of CAH1 were
fused to a promoterless arylsulfatase reporter gene (ARS). The upstream region from  651 to +41 relative to
the transcription start site was sufficient to regulate the expression
of ARS with kinetics similar to those of endogenous
CAH1. Deletion of the region between 651 and 294
resulted in a significant decrease in the level of arylsulfatase
activity expressed under low-CO2 conditions. The 543-bp
upstream region from 651 to 109, without any promoter elements,
CAAT-box, or TATA-box, could confer CO2 and light
responsiveness on the  2-tubulin minimal promoter. This 543-bp region was divided into two parts: a 358-bp silencer region from
651 to 294, which represses the minimal promoter activity under
high-CO2 conditions, and a 185-bp enhancer region from
293 to 109, which activates the promoter under low-CO2
conditions in the presence of light.
1
This work was supported by the Japanese Ministry
of Education, Science and Culture (grant nos. 09660357 and 10170219)
and by the Japan Society for the Promotion of Science (grant no.
JSPS-RFTF97R16001).
*
Corresponding author; e-mail fukuzawa{at}kais.kyoto-u.ac.jp; fax
81-75-753-6127.
© 1999 American Society of Plant Physiologists
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