Identification of oxygen dependent signalling pathways in the green alga Chlamydomonas reinhardtii by Digital Gene Expression Tag Profiling

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This project aims at identifying genes involved in oxygen (O2) signaling in the unicellular green alga Chlamydomonas reinhardtii, a well-studied reference alga whose genome has been sequenced. For this purpose, whole-genome transcription profiles of C. reinhardtii wild type cultures passing gradually into hypoxia are recorded using the next generation sequencing technology on the Illumina platform (RNA-seq). Gradual sampling allows identification of genes that are differentially expressed especially at the time-point when O2 starts to be limiting and whose protein products are therefore likely to play a role in O2-signalling. The original project plan has been revised to include several C. reinhardtii mutant strains deficient in the function of Crr1, a key transcriptional regulator essential for copper (Cu) and hypoxia signaling in C. reinhardtii. Very recent and yet unpublished results from both the Merchant (UCLA, USA) and Happe (RUB, Germany) groups indicate that Crr1 might play a much more important role in O2 dependent gene regulation than expected. Thus, the comparison of transcriptomes of wild type and crr1-mutant strains subjected to different O2 concentrations will allow me to gain more detailed insights into O2 signaling of C. reinhardtii. The extended project period will permit a deep analysis of sequence data by collaborating with local computational biologists.