Abstract
Most members of Siboglinidae (Annelida) harbor endosymbiotic bacteria that allow them to thrive in extreme environments such as hydrothermal vents, methane seeps, and whale bones. These symbioses are enabled by specialized hemoglobins (Hbs) that are able to bind hydrogen sulfide for transportation to their chemosynthetic endosymbionts. Sulfur-binding capabilities are hypothesized to be due to cysteine residues at key positions in both vascular and coelomic Hbs, especially in the A2 and B2 chains. Members of the genus Osedax, which live on whale bones, do not have chemosynthetic endosymbionts, but instead harbor heterotrophic bacteria capable of breaking down complex organic compounds. Although sulfur-binding capabilities are important in other siboglinids, we questioned whether Osedax retained these cysteine residues and the potential ability to bind hydrogen sulfide. To answer these questions, we used high-throughput DNA sequencing to isolate and analyze Hb sequences from 8 siboglinid lineages. For Osedax mucofloris, we recovered three (A1, A2, and B1) Hb chains, but the B2 chain was not identified. Hb sequences from gene subfamilies A2 and B2 were translated and aligned to determine conservation of cysteine residues at previously identified key positions. Hb linker sequences were also compared to determine similarity between Osedax and siboglinids/sulfur-tolerant annelids. For O. mucofloris, our results found conserved cysteines within the Hb A2 chain. This finding suggests that Hb in O. mucofloris has retained some capacity to bind hydrogen sulfide, likely due to the need to detoxify this chemical compound that is abundantly produced within whale bones.
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Acknowledgments
We are indebted to the late Christopher Schander for his contribution of Osedax mucofloris tissue for sequencing. We would like to thank the crews of the R/V Seward Johnson, R/V Aurelia, and R/V Håkons-Mosby for their assistance in procuring samples. We would also like to thank Amanda Shaver and Franziska Franke for preparation of cDNA for samples used in this study. Pamela M. Brannock, Kevin M. Kocot, and Nathan V. Whelan provided guidance and input on procedures used in this study and the writing of versions of this manuscript. This work was funded by National Science Foundation grants IOS-0843473 to K. M. H, S. R. S., and D. J. T., OCE-1155188 to K. M. H., DEB-1036537 to K. M. H. and S. R. S. It represents contributions #141 and #49 to the Auburn University (AU) Marine Biology Program and Molette Biology Laboratory for Environmental and Climate Change Studies, respectively.
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Genbank Accession Numbers KT166952–KT166980.
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Waits, D.S., Santos, S.R., Thornhill, D.J. et al. Evolution of Sulfur Binding by Hemoglobin in Siboglinidae (Annelida) with Special Reference to Bone-Eating Worms, Osedax . J Mol Evol 82, 219–229 (2016). https://doi.org/10.1007/s00239-016-9739-7
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DOI: https://doi.org/10.1007/s00239-016-9739-7