Subzero Temperature Adaptations Antifreeze Proteins

John G. Duman     Gillen Professor of Biological Sciences Ph.D., University of California, San Diego (Scripps)

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Research concerns the physiological and biochemical adaptations of poikilothermic organisms to subzero temperature. Most studies are with insects and plants, but other organisms (including spiders and other terrestrial invertebrates, fungi and bacteria) are also under investigation. These organisms adapt to subzero winter temperatures by either becoming freeze tolerant (able to survive extracellular freezing) or freeze avoiding.  Freeze avoiding species generally produce antifreezes, such as polyols (glycerol, etc) and/or antifreeze proteins. We have been particularly concerned with the antifreeze proteins and our studies have ranged from investigations of the function of the proteins, to hormonal and environmental cues controlling their production, to protein chemistry and molecular biology designed to determine the structure - antifreeze function relationships of the proteins.  Studies with freeze tolerant organisms have concentrated on ice nucleating proteins which function to induce ice formation in the extracellular fluid at high subzero temperature, and also on antifreeze proteins. In addition to functioning as antifreezes in freeze avoiding organisms, antifreeze proteins appear to function in certain freeze tolerant organisms as cryoprotectants to inhibit the damage resulting from freezing of body water. The mechanism of this process is under investigation.  Current ongoing research includes: (1) structure/function relationships of insect, plant and bacterial antifreeze proteins; (2) the cloning and expression of antifreeze protein genes; (3) potential cryoprotection mechanisms of antifreeze proteins; (4) applied studies on the potential uses of antifreeze proteins in agriculture (i.e., transgenic plants which produce insect antifreeze proteins) and for the cryopreservation of biological materials; and (5) studies of antifreeze proteins in Alaskan insects. However, within the broad theme of low temperature adaptations a wide variety of student projects may be accommodated. These may range from physiological ecology to protein biochemistry.

 

Selected Publications:

Olsen, T.M. and Duman, J.G. (1997) Maintenance of the supercooled state in overwintering Pyrochroid beetle larvae Dendroides canadensis: role of hemolymph ice nucleators and antifreeze proteins. J. Comp. Physiol. B, 167:105-113.

Olsen, T.M., Sass, S.J., Li, N. and Duman, J.G. (1998) Factors contributing to seasonal increases in inoculative freezing resistance in overwintering fire-colored beetle larvae Dendroides canadensis (Pyrochroidae). J. Exp. Biol. 201:1585-1594.

Duman, J.G., Parmalee, D., Goetz, F.W., Li, N., Wu, D.W., and Benjamin, T. (1998) Molecular characterization and sequencing of antifreeze proteins from larvae of the beetle Dendroides canadensis. J. Comp. Physiol. B, 168:225-232.

Li, N., Chibber, B.A.K., Castellino, F.J., and Duman, J.G. (1998) Mapping of disulfide bridges in antifreeze proteins from overwintering larvae of the beetle Dendroides canadensis. Biochemistry 37:6343-6350.

Li, N., Andorfer, C.A. and Duman, J.G. (1998) Enhancement of insect antifreezeprotein activity by low molecular weight solutes. J. Exp. Biol. 201:2243-2251.

Andorfer, C.A. and Duman, J.G. Isolation and characterization of cDNA clones encoding antifreeze proteins of the Pyrochroid beetle Dendroides canadensis . (2000). J. Insect. Physiol. 46:365-372.

Newton, S.S. and Duman, J.G. (2000) An osmotin-like cryoprotective protein from the bittersweet nightshade Solanum dulcamara . Plant Mol. Biol. 44:581-589.

Duman, J.G. (2001) Antifreeze and ice nucleator proteins in terrestrial arthropods. Ann. Rev. Physiol. 63:327-357.

Huang, T. and Duman, J.G. (2002) Cloning and characterization of a thermal hysteresis (antifreeze) protein with DNA-binding activity from winter bittersweet nightshade, Solanum dulcamara. Plant Mol. Biol. 48, 339-350.

Duman, J.G. (2002) The inhibition of ice nucleators by insect antifreeze proteins is enhanced by glycerol and citrate.  J. Comp. Physiol. B. 172, 163-168.

Duman, J.G. and Serianni, A.S. (2002) The role of endogenous antifreeze protein enhancers in the hemolymph thermal hysteresis activity of the beetle Dendroides canadensis. J. Insect Physiol. 48, 103-111.

Huang, Tao, Nicodemus, Jessie, Zarka, Daniel G., Thomashow, Michael F., Wisniewski, Michael and

Duman, John G. (2002).  Expression of an Insect (Dendroides canadensis) Antifreeze Protein in Arabidopsis thaliana Results in a Decrease in Plant Freezing Temperature. Plant Molecular Biology, 50:333-344.

Duman, J. G., Verleye, D. and Li, N. (2002). Site specific forms of antifreeze protein in the beetle Dendroides canadensis . J. Comp. Physiol. B., 172:547-552.

Duman, J.G., Bennett V., Sformo T., Hochstrasser R., and Barnes B.M. (2004) Antifreeze proteins in Alaskan insects and spiders. J. INsect Physiol. 50: 259-266.

Wang, L. and Duman, J.G. (2005) Antifreeze proteins of the beetle Dendroides canadensis enhance one antoher's activities.  Biolchemistry 44: 10305-10312.

Bennett, V.A., Sformo, T., Walters, K., Toien, O., Jeannet, K., Hochstrasser, R., Pan, Q., Serianni, A.S., Barnes, B.M., and Duman, J.G. (2005) Comparative overwintering physiology of Alaska and Indiana populations of the beetle Cucujus clavipes (Fabricus): Roles of antifreeze proteins, polyols, dehydration, and diapause. Journal of Experimental Biology 208: 4467-4477.

Wang, L., and Duman, J.G. (2006) A thaumatin-like protein from larvae of the beetle Dendroides canadensis enhances the activity of antifreeze proteins. Biochemistry 45: 1278-1284.