NSF Org:	OPP Office of Polar Programs (OPP)
Recipient:	UNIVERSITY OF ALASKA FAIRBANKS
Initial Amendment Date:	May 14, 2004
Latest Amendment Date:	May 14, 2004
Award Number:	0352919
Award Instrument:	Standard Grant
Program Manager:	Jane V. Dionne OPP  Office of Polar Programs (OPP) GEO  Directorate for Geosciences
Start Date:	June 1, 2004
End Date:	May 31, 2006 (Estimated)
Total Intended Award Amount:	$114,964.00
Total Awarded Amount to Date:	$114,964.00
Funds Obligated to Date:	FY 2004 = $114,964.00
History of Investigator:	Brian Barnes (Principal Investigator) bmbarnes@alaska.edu
Recipient Sponsored Research Office:	University of Alaska Fairbanks Campus 2145 N TANANA LOOP FAIRBANKS AK  US  99775-0001 (907)474-7301
Sponsor Congressional District:	00
Primary Place of Performance:	University of Alaska Fairbanks Campus 2145 N TANANA LOOP FAIRBANKS AK  US  99775-0001
Primary Place of Performance Congressional District:	00
Unique Entity Identifier (UEI):	FDLEQSJ8FF63
Parent UEI:
NSF Program(s):	ARC Rsch Support & Logistics, ANS-Arctic Natural Sciences
Primary Program Source:	0100CYXXDB NSF RESEARCH & RELATED ACTIVIT
Program Reference Code(s):	OTHR, 1079, 0000
Program Element Code(s):	520500, 528000
Award Agency Code:	4900
Fund Agency Code:	4900
Assistance Listing Number(s):	47.078

ABSTRACT

This is a collaborative proposal by Principal Investigators at the University of Alaska-Fairbanks and Purdue University. Antifreeze proteins (AFPs) are found in a number of diverse organisms including fish, insects, spiders, mites, plant, and bacteria. While there is tremendous structural variation among these AFPs they all appear to lower the freezing point of water by a similar non-colligative mechanism. However, the insect AFPs are, by far, the most active, generally functioning to prevent freezing in freeze susceptible insects by inhibiting ice nucleating agents and innoculative freezing across the cuticle. However, a few AFP-producing insects are freeze tolerant and the function of AFPs in these is unknown. Two insect AFPs have been characterized. One type has been described from two species of beetles, but there is variation in sequence, etc., both within and between the species. Until recent research identified 18 species with AFPs, these proteins had not been reported in Alaskan or arctic insects. Certain of these have been chosen for further study, presenting the opportunity to extend understanding of insect AFPs to extremely cold tolerant species, and in the case of the beetle Cucujus clavipes ,to compare populations from arctic Alaska to those from Indiana. The mean winter supercooling point of Alaskan Cucujus larvae are -42oC, with some individuals supercooling to -57oC, while that of Indiana larvae is -24oC. AFP activity in Alaskan Cucujus is, by far, the highest ever described, even though the AFPs are similar to those of other beetles. Understanding the structure/function relationships of the Cucujus AFPs, and the enhancers (other proteins and polyols) which increase their activity, is of prime concern. In addition to AFPs, a suite of other overwintering adaptations (extreme dehydration to 1/3 normal body water, multimolar glycerol concentrations, diapause) contribute to the low supercooling points of Alaskan Cucujus. These are being studied, and using microarray technology and other means to monitor molecular markers, the Principal Investigators will attempt to understand the integration of these adaptations. Additional investigations involve the structure/function relationships of three other Alaskan beetle AFPs, and that of the insect type of AFP. Two of the beetle species are freeze tolerant and this is the first study of AFPs from freeze tolerant insects. The Principal Investigators will continue to screen additional species for the presence of AFPs (to identify interesting candidates for future study), and to monitor microhabitat temperatures and overwintering mortality of select species. The broader impact of this study concerns its effects on 1) biological education and 2) applications of AFPs in science and technology. Post-docs, PhD and undergraduate students, and high school teachers will work on this project. The Principal Investigators believe that the integrated training (in modern molecular techniques such as microarray, field biology, physiology of cold tolerance, protein biochemistry, etc.) at they will receive provides critical preparation for success in today's highly
integrative and changing research environment. This will provide a direct benefit to teaching in biology. In addition to research papers, the Principal Investigators will incorporate these studies in review articles, symposium presentations. Also, as they are the most active AFPs known, insect AFPs (especially from Alaskan beetles) have tremendous potential for applied studies. For example, they have generated transgenic plants producing insect AFPs which lower the plant freezing temperature. Similar work using Alaskan beetle AFPs should be even more productive.

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