Cadmium- and iron-stress-inducible gene expression in the green alga Chlamydomonas reinhardtii: evidence for H43 protein function in iron assimilation

Abstract.

Early transcriptional responses of a cell wall-deficient mutant of the green alga Chlamydomonas reinhardtii to heavy-metal stress have been investigated using the method of mRNA differential display. We have identified, sequenced, and quantified the induction of a number of transcripts that are up-regulated by a brief (2-h) exposure to 25 µm cadmium chloride, including one transcript which is also highly responsive to iron (Fe) deficiency. These transcripts represent both nuclear- and chloroplast-encoded genes, and include both novel genes and genes with known or suspected functions. Among these is a gene with significant homology to HCR1, a high-CO₂- and Fe-deficiency-inducible gene from Chlorococcum littorale. We further characterized the regulation of the HCR1-like gene (H43) and found that this transcript is also induced by Fe-depletion of the medium. Heterologous expression of H43 in the Fe-uptake mutant fet3fet4 of Saccharomyces cerevisiae resulted in partial suppression of the slow-growth phenotype of this mutant in minimal medium, and resulted in a 2-fold increase in Fe accumulation per cell. Our results demonstrate the utility of Chlamydomonas cw ^– strains for functional genomics studies of metal stress. The magnitudes of induction and functional analyses suggest possible utility for these genes in the study of metal stress sensing in green plants and development of novel Fe acquisition and phytoremediation strategies.