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Characterization and expression of genes from the RubisCO gene cluster of the chemoautotrophic symbiont of Solemya velum: cbbLSQO

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Abstract

Chemoautotrophic endosymbionts residing in Solemya velum gills provide this shallow water clam with most of its nutritional requirements. The cbb gene cluster of the S. velum symbiont, including cbbL and cbbS, which encode the large and small subunits of the carbon-fixing enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO), was cloned and expressed in Escherichia coli. The recombinant RubisCO had a high specific activity, ∼3 μmol min−1 mg protein −1, and a K CO2 of 40.3 μM. Based on sequence identity and phylogenetic analyses, these genes encode a form IA RubisCO, both subunits of which are closely related to those of the symbiont of the deep-sea hydrothermal vent gastropod Alviniconcha hessleri and the photosynthetic bacterium Allochromatium vinosum. In the cbb gene cluster of the S. velum symbiont, the cbbLS genes were followed by cbbQ and cbbO, which are found in some but not all cbb gene clusters and whose products are implicated in enhancing RubisCO activity post-translationally. cbbQ shares sequence similarity with nirQ and norQ, found in denitrification clusters of Pseudomonas stutzeri and Paracoccus denitrificans. The 3′ region of cbbO from the S. velum symbiont, like that of the three other known cbbO genes, shares similarity to the 3′ region of norD in the denitrification cluster. This is the first study to explore the cbb gene structure for a chemoautotrophic endosymbiont, which is critical both as an initial step in evaluating cbb operon structure in chemoautotrophic endosymbionts and in understanding the patterns and forces governing RubisCO evolution and physiology.

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Acknowledgments

We would like to thank Jonathan Robinson, Meredith Hullar, and Adam McCoy for helpful discussions, Martin Polz for help with automated sequencing, and Zoe McKiness for assistance with phylogenetic analyses. We thank Robert Fisher for sending plasmids and George Lorimer for RubisCO antisera. This work was supported by NSF grants OCE-9504257 and OCE-0002460, and by NASA grant NAG5-10906.

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  1. Julie Schwedock

    Present address: Genomic Profiling Systems, Bedford, MA, 01730, USA

  2. Kathleen M. Scott

    Present address: Department of Biology, University of South Florida, Tampa, FL, 33620, USA

  3. Daniel L. Distel

    Present address: Department of Biochemistry, Microbiology, and Molecular Biology, University of Maine, Orono, ME, 04469, USA

Authors and Affiliations

  1. Department of Organismic and Evolutionary Biology, Harvard University, 16 Divinity Avenue, Cambridge, MA, 02138, USA

    Julie Schwedock, Tara L. Harmer, Kathleen M. Scott, Harm J. Hektor, Angelica P. Seitz, Matthew C. Fontana, Daniel L. Distel & Colleen M. Cavanaugh

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  1. Julie Schwedock
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Correspondence to Colleen M. Cavanaugh.

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Schwedock, J., Harmer, T.L., Scott, K.M. et al. Characterization and expression of genes from the RubisCO gene cluster of the chemoautotrophic symbiont of Solemya velum: cbbLSQO . Arch Microbiol 182, 18–29 (2004). https://doi.org/10.1007/s00203-004-0689-x

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