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Evolution of Hyperactive, Repetitive Antifreeze Proteins in Beetles

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Abstract

Some organisms that experience subzero temperatures, such as insects, fish, bacteria, and plants, synthesize antifreeze proteins (AFPs) that adsorb to surfaces of nascent ice crystals and inhibit their growth. Although some AFPs are globular and nonrepetitive, the majority are repetitive in both sequence and structure. In addition, they are frequently encoded by tandemly arrayed, multigene families. AFP isoforms from the mealworm beetle, Tenebrio molitor, are extremely potent and inhibit ice growth at temperatures below −5°C. They contain a 12-amino acid repeat with the sequence TCTxSxxCxxAx, each of which makes up one coil of the β-helix structure. TxT motifs are arrayed to form the ice-binding surface in all three known insect AFPs: the homologous AFPs from the two beetles, T. molitor and Dendroides canadensis, and the nonhomologous AFP from the spruce budworm, Choristoneura fumiferana. In this study, we have obtained the cDNA and genomic sequences of additional T. molitor isoforms. They show variation in the number of repeats (from 6 to 10) which can largely be explained by recombination at various TCT motifs. In addition, phylogenetic comparison of the AFPs from the two beetles suggests that gene loss and amplification may have occurred after the divergence of these species. In contrast to a previous study suggesting that T. molitor genes have undergone positive Darwinian selection (selection for heterogeneity), we propose that the higher than expected ratio of nonsynonymous-to-synonymous substitutions might result from selection for higher AT content in the third codon position.

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Acknowledgments

CHIR and NSERC (Canada) are acknowledged for grants to P.L.D and V.K.W., respectively. The authors thank Dr. D. Forsdyke for comments on the manuscript. The comments made by the reviewers were invaluable and helped us to dramatically improve our analysis and interpretation of the results.

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Authors and Affiliations

  1. Department of Biochemistry, Queen’s University, Kingston, Ontario, K7L 3N6, Canada

    Laurie A. Graham & Peter L. Davies

  2. Department of Biology, Queen’s University, Kingston, Ontario, K7L 3N6, Canada

    Wensheng Qin, Stephen C. Lougheed, Peter L. Davies & Virginia K. Walker

  3. Protein Function Discovery Group, Queen’s University, Kingston, Ontario, K7L 3N6, Canada

    Peter L. Davies & Virginia K. Walker

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  1. Laurie A. Graham
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  2. Wensheng Qin
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  3. Stephen C. Lougheed
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  4. Peter L. Davies
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  5. Virginia K. Walker
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Correspondence to Virginia K. Walker.

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[Reviewing Editor: Dr. John Oakeshott]

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Graham, L.A., Qin, W., Lougheed, S.C. et al. Evolution of Hyperactive, Repetitive Antifreeze Proteins in Beetles. J Mol Evol 64, 387–398 (2007). https://doi.org/10.1007/s00239-005-0256-3

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  • DOI: https://doi.org/10.1007/s00239-005-0256-3

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