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Eukaryotic Cell, October 2003, p. 995-1002, Vol. 2, No. 5
1535-9778/03/$08.00+0     DOI: 10.1128/EC.2.5.995-1002.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Copper Response Element and Crr1-Dependent Ni2+-Responsive Promoter for Induced, Reversible Gene Expression in Chlamydomonas reinhardtii

Jeanette M. Quinn,{dagger} Janette Kropat, and Sabeeha Merchant*

Department of Chemistry and Biochemistry, University of California at Los Angeles, Los Angeles, California 90095-1569

Received 18 June 2003/ Accepted 8 July 2003

The Cpx1 and Cyc6 genes of Chlamydomonas reinhardtii are activated in copper-deficient cells via a signal transduction pathway that requires copper response elements (CuREs) and a copper response regulator defined by the CRR1 locus. The two genes can also be activated by provision of nickel or cobalt ions in the medium. The response to nickel ions requires at least one CuRE and also CRR1 function, suggesting that nickel interferes with a component in the nutritional copper signal transduction pathway. Nickel does not act by preventing copper uptake/utilization because (i) holoplastocyanin formation is unaffected in Ni2+-treated cells and (ii) provision of excess copper cannot reverse the Ni-dependent activation of the target genes. The CuRE is sufficient for conferring Ni-responsive expression to a reporter gene, which suggests that the system has practical application as a vehicle for inducible gene expression. The inducer can be removed either by replacing the medium or by chelating the inducer with excess EDTA, either of which treatments reverses the activation of the target genes.

* Corresponding author. Mailing address: Department of Chemistry and Biochemistry, UCLA, Box 951569, Los Angeles, CA 90095-1569. Phone: (310) 825-8300. Fax: (310) 206-1035. E-mail: merchant{at}chem.ucla.edu.

{dagger} Present address: Stratagene, La Jolla, Calif.

Eukaryotic Cell, October 2003, p. 995-1002, Vol. 2, No. 5
1535-9778/03/$08.00+0     DOI: 10.1128/EC.2.5.995-1002.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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