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Molecular and biochemical analysis of the α class carbonic anhydrases in Caenorhabditis elegans

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Abstract

In this study, in silico analysis of the Caenorhabditis elegans genome revealed six genes (cah-1, cah-2, cah-3, cah-4, cah-5, and cah-6) possibly encoding α class CAs (carbonic anhydrase). Real-time RT-PCR analysis revealed the temporal expression pattern of each gene, as well as changes in expression levels under different atmospheric conditions (stress). Cah-3 and cah-4 showed the highest levels of transcript accumulation, while most genes responded to the stress conditions. Yeast complementation showed that cah-3 was able to complement the function of Saccharomyces cerevisiae CA (NCE103) in vivo. Recombinant CAH-3, CAH-4a and CAH-5 enzymes, expressed in Escherichia coli were used for in vitro measurement of CA activity. However, in vitro activity was only detectable for CAH-4a. RNAi by feeding was performed on wild-type C. elegans for all genes. The worms were examined for a visible phenotype under normal and stress conditions (pH, CO2/O2). Silencing cah-3 and cah-4 may reduce the life-span of the worms (at 22°C).

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Acknowledgments

We thank Prof. Y. Kohara from the Kohara Laboratory (NIG, Japan) for providing the cDNA clones yk713, yk443, yk1011 and yk794. We also thank Dr. N. Tavernarakis from the Tavernarakis Lab (IMBB, Greece) for his assistance and for providing the C. elegans strain N2, the E. coli strains OP50 and HT115 and the pL4440 vector.

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  1. Department of Biotechnology, Agricultural University of Athens, Iera Odos 75, 11855, Athens, Greece

    Michael K. Fasseas, Daniela Tsikou, Emmanouil Flemetakis & Panagiotis Katinakis

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  1. Michael K. Fasseas
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  2. Daniela Tsikou
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  3. Emmanouil Flemetakis
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  4. Panagiotis Katinakis
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Correspondence to Panagiotis Katinakis.

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Fasseas, M.K., Tsikou, D., Flemetakis, E. et al. Molecular and biochemical analysis of the α class carbonic anhydrases in Caenorhabditis elegans . Mol Biol Rep 38, 1777–1785 (2011). https://doi.org/10.1007/s11033-010-0292-y

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  • DOI: https://doi.org/10.1007/s11033-010-0292-y

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