Characterization of a novel candidate gene involved in the anaerobic response of Chlamydomonas reinhardtii as revealed by comparative proteomics

Mia Terashima, Michael Specht, Irina Tolstygina, Bianca Naumann, and Michael Hippler

Department of Biology, Institute of Plant Biochemistry and Biotechnology, University of Münster, Hindenburgplatz 55, 48143 Münster, Germany

Under anaerobic conditions, C. reinhardtii can produce H2 catalyzed by the hydrogenase. In order to better understand the proteome of C. reinhardtii under hydrogen-producing conditions, chloroplasts and mitochondria from aerobic and anaerobic (induced by 8 hours of argon bubbling) conditions were isolated and analyzed using comparative proteomics. Protein expressions between the two conditions were compared qualitatively and semi-quantitatively, allowing 606 proteins to be localized to the chloroplast (Terashima et al., 2010). In addition, quantitative analyses were performed with the chloroplast-localized proteins using stable isotopic labeling of amino acids (13C6 Arginine/12C6 Arginine in an Arg auxotrophic strain). The in-house developed automated quantitation program, qTrace, was used for high-throughput quantitation. Quantitative data showed TEF7, a green algae-specific protein of unknown function, to be induced under anaerobic conditions. TEF7 has been characterized through the development of several ami-RNA lines and biochemical analyses in order to further understand the possible role of this protein in the anaerobic response of C. reinhardtii. A model for the role of this protein based on current findings will be presented. Terashima, M., Specht, M., Naumann, B. and Hippler, M. (2010) Characterizing the anaerobic response of Chlamydomonas reinhardtii by quantitative proteomics. Mol Cell Proteomics, in press.

e-mail address of presenting author: mtera_01@uni-muenster.de