Cloning and characterization of LRG5, a gene involved in blue light signaling in Chlamydomonas gametogenesis

Abstract

The Chlamydomonas reinhardtii mutant Irg2 exhibits a partial defect in the blue-light activated signal transduction chain that controls gametogenesis. By genomic complementation, a cosmid clone that corrects the Irg2 phenotype was isolated. The smallest fragment of this cosmid that gave complementation was used in RFLP analysis, which revealed that the cloned gene was not linked to the LRG2 locus. The sequence of the genomic clone and a cDNA of this gene, which we named LRG5, was determined. Although no significant homology with sequences in the databases was found, the nuclear localization signal and the presence of negatively and positively charged domains suggests a function for the LRG5 protein in the regulation of transcription. The correction of the Irg2 phenotype is probably caused by overexpression of the LRG5 gene in the transformants, as these expressed a level of LRG5 approximately twofold higher than wild-type or mutant strains. Southern blot analysis provided evidence for the existence of homologous sequences in other green algae as well as in higher plants, suggesting a conserved function for the LRG5 protein.