Abstract
Hemocyanin is a copper-containing protein that transports O2 in the hemolymph of many arthropod species. Within the crustaceans, hemocyanin appeared to be restricted to Malacostraca but has recently been identified in Remipedia. Here, we report the occurrence of hemocyanin in ostracods, indicating that this respiratory protein is more widespread within crustaceans than previously thought. By analyses of expressed sequence tags and by RT-PCR, we obtained four full length and nine partial hemocyanin sequences from six of ten investigated ostracod species. Hemocyanin was identified in Myodocopida (Actinoseta jonesi, Cypridininae sp., Euphilomedes morini, Skogsbergia lerneri, Vargula tsujii) and Platycopida (Cytherelloidea californica) but not in Podocopida. We found no evidence for the presence of hemoglobin in any of these ostracod species. Like in other arthropods, we identified multiple hemocyanin subunits (up to six) to occur in a single ostracod species. Bayesian phylogenetic analyses showed that ostracod hemocyanin subunit diversity evolved independently from that of other crustaceans. Ostracod hemocyanin subunits were found paraphyletic, with myodocopid and platycopid subunits forming distinct clades within those of the crustaceans. This pattern suggests that ostracod hemocyanins originated from distinct subunits in the pancrustacean stemline.
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Abbreviations
- BPP:
-
Bayesian posterior probability
- Hc:
-
Hemocyanin
- MYA:
-
Million years ago
References
Abascal F, Zardoya R, Posada D (2005) ProtTest: selection of best-fit models of protein evolution. Bioinformatics 21:2104–2105
Abe K, Vannier J (1995) Functional morphology and significance of the circulatory system of Ostracoda, exemplified by Vargula hilgendorfii (Myodocopida). Mar Biol 124:51–58
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
Bowman TE, Abele LG (1982) Classification of the recent crustacea. In: Abele LG (ed) The Biology of Crustacea, vol 1., Systematics, the Fossil Record and BiogeographyAcademic Press, New York, London, pp 1–27
Burmester T (1999) Identification, molecular cloning and phylogenetic analysis of a non-respiratory pseudo-hemocyanin of Homarus americanus. J Biol Chem 274:13217–13222
Burmester T (2001) Molecular evolution of the arthropod hemocyanin superfamily. Mol Biol Evol 18:184–195
Burmester T (2002) Origin and evolution of arthropod hemocyanins and related proteins. J Comp Physiol B 172:95–107
Corbari L, Carbonel P, Massabuau JC (2004a) How a low tissue O2 strategy could be conserved in early crustaceans: the example of the podocopid ostracods. J Exp Biol 207:4415–4425
Corbari L, Carbonel P, Massabuau JC (2004b) The early life history of tissue oxygenation in crustaceans: the strategy of the myodocopid ostracod Cylindroleberis mariae. J Exp Biol 208:661–670
Dewilde S, Ebner B, Vinck E, Gilany K, Hankeln T, Burmester T, Kreiling J, Reinisch C, Vanfleteren JR, Kiger L, Marden MC, Hundahl C, Fago A, van Doorslaer S, Moens L (2006) The nerve hemoglobin of the bivalve mollusc Spisula solidissima, molecular cloning, ligand binding studies, and phylogenetic analysis. J Biol Chem 281:5364–5372
Ertas B, von Reumont BJ, Wägele JW, Misof B, Burmester T (2009) Hemocyanin suggests a close relationship of Remipedia and Hexapoda. Mol Biol Evol 26:2711–2718
Ertas B, Kiger L, Blank M, Marden MC, Burmester T (2011) A membrane-bound hemoglobin from gills of the Green Shore Crab Carcinus maenas. J Biol Chem 286:3185–3193
Fox HM (1957) Haemoglobin in the Crustacea. Nature 179:148
Gaykema WPJ, Hol WGJ, Vereifken JM, Soeter NM, Bak HJ, Beintema JJ (1984) 3.2 Å structure of the copper-containing oxygen-carrying protein Panulirus interruptus hemocyanin. Nature 309:23–29
Herberts C, de Frescheville J (1981) Occurrence of hemocyanin in. the rhizocephalan crustacea Sacculina carcini Thompson. Comp Biochem Physiol B 70:657–659
Horne DJ, Cohen A, Martens K (2002) Taxonomy, morphology and biology of quaternary and living ostracoda. In: Holmes JA, Chivas AR (eds) The ostracoda: applications in quaternary research, vol 131. Geophysical Monograph American Geophysical Union, Washington, DC, pp 5–36
Horne DJ, Schön I, Smith RJ, Martens K (2005) What are Ostracoda? A cladistics analysis of the extant superfamilies of the subclasses Myodocopa and Podocopa (Crustacea, Ostracoda). In: Koenemann S, Jenner RA (eds) Crustacea and Arthropod Relationships. CRC Press, Boca Raton (FL), pp 249–273
Huelsenbeck JP, Ronquist F (2001) MRBAYES: bayesian inference of phylogenetic trees. Bioinformatics 17:754–755
Ikeya N, Tsukagoshi A, Horne DJ (2005) Preface: The phylogeny, fossil record and ecological diversity of ostracod crustaceans. Hydrobiologia 538:vii–xiii. In: Ikeya N, Tsukagoshi A, Horne D J (eds.): Evolution and Diversity of Ostracoda. 14th International Symposium on Ostracoda (ISO 2001), Shizuoka, Japan
Katoh K, Kumal K, Tohl H, Miyata T (2005) MAFFT version 5: improvement in accuracy of multiple sequence alignment. Nucleic Acids Res 33:511–518
Keyser D (1990) Morphological changes and function of the inner lamella layer of podocopid Ostracoda. In: Whatley R, Maybury C (eds) Ostracoda and Global Events. Chapman and Hall, London, pp 401–410
Kusche K, Burmester T (2001) Diplopod hemocyanin sequence and the phylogenetic position of the Myriapoda. Mol Biol Evol 18:1566–1573
Kusche K, Ruhberg H, Burmester T (2002) A hemocyanin from the Onychophora and the emergence of respiratory proteins. Proc Natl Acad Sci USA 99:10545–10548
Kusche K, Hembach A, Milke C, Burmester T (2003) Molecular characterisation and evolution of the hemocyanin from the European spiny lobster, Palinurus elephas. J Comp Physiol B 173:319–325
Linzen B, Soeter NM, Riggs AF, Schneider HJ, Schartau W, Moore MD, Yokata E, Behrens PQ, Nakashima H, Takagi T, Nemoto T, Vereijken JM, Bak HJ, Beintema JJ, Volbeda A, Gaykema WPJ, Hol WGJ (1985) The structure of arthropod hemocyanins. Science 229:519–524
Mangum CP (1985) Oxygen transport in invertebrates. Am J Physiol 248:505–514
Markl J (1986) Evolution and function of structurally diverse subunits in the respiratory protein hemocyanin from arthropods. Biol Bull 171:90–115
Markl J, Decker H (1992) Molecular structure of the arthropod hemocyanins. In: Mangum CP (ed) Advances in Comparative and Environmental Physiology, vol 13., pp 325–376
Markl J, Stocker W, Runzler R, Precht E (1986) Immunological correspondences between the hemocyanin subunits of 86 arthropods: evolution of a multigene protein family. In: Linzen B (ed) Invertebrate Oxygen Carriers. Springer-Verlag, Heidelberg, pp 281–292
Martens K, Schön I, Meisch C, Horne DJ (2008) Global diversity of ostracods (Ostracoda, Crustacea) in freshwater. Hydrobiologia 595:185–193
Marxen JC, Pick C, Kwiatkowski M, Burmester T (2013) Molecular characterization and evolution of hemocyanin from the two Freshwater shrimps Caridina multidentata (Stimpson, 1860) and Atyopsis moluccensis (De Haan, 1849). J Comp Physiol B 183:623–624
Nicholas KB, Nicholas HBJ, Deerfield DWI (1997) GeneDoc: analysis and visualization of genetic variation. EMBnet NEWS 4:14
Oakley TH, Wolfe JM, Lindgren AR, Zaharoff AK (2013) Phylotranscriptomics to bring the understudied into the fold: monophyletic ostracoda, fossil placement, and pancrustacean phylogeny. Mol Biol Evol 30:215–233
Petersen TN, Brunak S, von Heijne G, Nielsen H (2011) SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat Methods 8:785–786
Pick C, Schneuer M, Burmester T (2009) The occurrence of hemocyanin in Hexapoda. FEBS J 276:1930–1941
Regier JC, Shultz JW, Zwick A, Hussey A, Ball B, Wetzer R, Martin JW, Cunningham CW (2010) Arthropod relationships revealed by phylogenomic analysis of nuclear protein-coding sequences. Nature 463:1079–1083
Scherbaum S, Beyhan E, Gebauer W, Burmester T (2010) Characterization of hemocyanin from the peacock mantis shrimp Odontodactylus scyllarus (Malacostraca: hoplocarida). J Comp Physiol B 180:1235–1245
Smith AJ, Horne DJ (2002) Ecology of marine, marginal marine and nonmarine ostracodes. In: Holmes JA, Chivas AR (eds) The Ostracoda: applications in quaternary research, vol 131. Geophysical Monograph American Geophysical Union, Washington, DC, pp 37–64
Stamatakis A (2006) RAxML-VI-HPC: maximum Likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics 22:2688–2690
Terwilliger NB, Dangott L, Ryan M (1999) Cryptocyanin, a crustacean molting protein: evolutionary link with arthropod hemocyanins and insect hexamerins. Proc Natl Acad Sci USA 96:2013–2018
Tinn O, Oakley TH (2008) Erratic rates of molecular evolution and incongruence of fossil and molecular divergence time estimates in Ostracoda (Crustacea). Mol Phylogenet Evol 48:157–167
van Holde KE, Miller KI (1995) Hemocyanins. Adv Prot Chem 47:1–81
Vannier J, Abe K (1995) Size, body plan and respiration in the Ostracoda. Palaeontology 38:843–873
Volbeda A, Hol WGJ (1989) Crystal structure of hexameric haemocyanin from Panulirus interruptus refined at 3.2 Å resolution. J Mol Biol 209:249–279
von Reumont BM, Jenner RA, Wills MA, Dell’Ampio E, Pass G, Ebersberger I, Meyer B, Koenemann S, Iliffe TM, Stamatakis A, Niehuis O, Meusemann K, Misof B (2012) Pancrustacean phylogeny in the light of new phylogenomic data: support for Remipedia as the possible sister group of Hexapoda. Mol Biol Evol 29:1031–1045
Weber RE, Vinogradov SN (2001) Nonvertebrate Hemoglobins: functions and Molecular Adaptations. Physiol Rev 81:569–628
Whelan S, Goldman N (2001) A general empirical model of protein evolution derived from multiple protein families using a Maximum-Likelihood approach. Mol Biol Evol 18:691–699
Acknowledgments
We thank Janus Borner for access to the program Primerlyze, Dietmar Keyser for his help with species determination, and Elizabeth Torres for her numerous attempts to collect S. lerneri. This work was supported by the Deutsche Forschungsgemeinschaft (BU 956/14) to TB and by the National Science Foundation (DEB-1146337) to THO.
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Marxen, J.C., Pick, C., Oakley, T.H. et al. Occurrence of Hemocyanin in Ostracod Crustaceans. J Mol Evol 79, 3–11 (2014). https://doi.org/10.1007/s00239-014-9636-x
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DOI: https://doi.org/10.1007/s00239-014-9636-x