Subzero
Temperature Adaptations, Antifreeze Proteins
| John
G. Duman
Gillen Professor of Biological Sciences
Ph.D.,
University of California, San Diego (Scripps Institution of Oceanography) |
|
Research
concerns the physiological and biochemical adaptations
of poikilothermic organisms to subzero temperature. Most
studies are on insects, but other organisms
(including plants, spiders and other terrestrial invertebrates) 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 (AFPs). We have been
particularly concerned with the AFPs, 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 their AFPs. Antifreeze proteins
function in certain freeze tolerant organisms
as cryoprotectants to inhibit the damage resulting from
freezing of body water. The mechanisms of this process
are under investigation, but include recrystallization inhibition (control of ice crystal size) and inhibition of the generally lethal spread of extracellular ice into the cytoplasm. Recently, a new type of thermal hysteresis factor/antifreeze that is a glycolipid has been characterized. The antifreeze glycolipid has levels of antifreeze activity comparable to insect AFPs and is associated mainly with cell membranes where it may inhibit intracellular ice formation. Current research
includes: (1) structure/function relationships of insect and
plant antifreeze proteins and glycolipids; (2) the cloning
and expression of antifreeze protein genes; (3) cryoprotection mechanisms of antifreeze proteins and glycolipids in freeze tolerance; (4)
applied studies on (a) the potential uses of antifreeze proteins
and glycolipids in agriculture (i.e., transgenic plants which produce
insect antifreeze proteins) and (b) the cryopreservation
of biomedically important tissues and organs; and (5) studies of antifreeze
proteins and related adaptations in Alaskan insects, including proteomic and other studies of how the beetle
Cucujus clavipes deep supercools to -100 degrees C. Within the broad
theme of low temperature adaptations a wide variety of
student projects, ranging from physiological ecology to protein biochemistry, can be accommodated.
Selected
Publications:
Duman, J.G. (2001) Antifreeze and ice nucleator proteins in terrestrial arthropods. Annual Review of Physiology 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 Molecular Biology 48:339-350.
Duman, J.G. (2002) The inhibition of ice nucleators by insect antifreeze proteins is enhanced by glycerol and citrate. Journal of Comparative Physiology B. 172:163-168.
Huang, T., Nicodemus, J., Zarka, D.G., Thomashow,M.F. and Duman ,J.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 Proteins in the Beetle Dendroides canadensis. Journal of Comparative Physiology 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. Journal of Insect Physiology 50: 259-266.
Wang, L. and Duman, J.G. (2005) Antifreeze proteins of the beetle Dendroides canadensis enhance one another's activities. Biochemistry 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.
Nicodemus, J., O’Tousa, J.E., and Duman, J.G. (2006) Expression of a beetle, Dendroides canadensis, antifreeze protein in Drosophila melanogaster. Journal of Insect Physiology 52: 888-896.
Walters, K.R., Sformo, T., Barnes, B.M., and Duman, J.G. (2009) Freeze tolerance of an Arctic Alaska stonefly. Journal of Experimental Biology 212: 305-312.
Sformo, T., Kohl, F., McIntyre, J., Kerr, P., Duman, J. G., and Barnes, B. M. (2009) Simultaneous freeze tolerance and avoidance in individual fungus gnats, Exechia migratoria. Journal of Comparative Physiology B 179: 897-902.
Walters, K. R., Pan, Q., Serianni, A. S., and Duman, J., G. (2009) Cryoprotectant biosynthesis and selective accumulation of threitol in the freeze tolerant beetle, Upis ceramboides. Journal of Biological Chemistry 284: 16,822-.
Walters, K.R., Serianni, A.S., Sformo, T., Barnes, B.M., Duman, J.G. (2009) A novel thermal hysteresis-producing xylomannan antifreeze in a freeze tolerant Alaskan beetle. Proceedings of the National Academy of Science (in press).
Duman, J. G., Walters, K. R., Sformo, T., Carrasco, M. A., Nickell, P. and Barnes, B. M. (2010) Antifreeze and ice nucleator proteins. In “Low Temperature Biology of Insects” (D. Denlinger and R. E. Lee, eds) Cambridge University Press, Cambridge, UK. pp 59-90.
Lin, X., O’Tousa, J.E., and Duman, J.G. (2010) Expression of two self-enhancing antifreeze proteins from the beetle Dendroides canadensis in Drosophila melanogaster. Journal of Insect Physiology (in press).
Sformo T, Walters K, Jeannet K, McIntyre J, Wowk B, Fahy G, Barnes BM, Duman JG. (2010) Deep Supercooling, vitrification, and limited survival to –100oC in the Alaskan beetle larvae Cucujus clavipes puniceus (Coleoptera: Cucujuidae). Journal of Experimental Biology (in press).
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