Protein Folding and Function at Cold Temperature: Co-Evolution of the Chaperonin CCT and Tubulins from Antarctic Fishes
Since the advent of Antarctic continental glaciation, the opening of the Drake Passage between South America and the Antarctic Peninsula, and the onset of cooling of the Southern Ocean ~38-25 million years ago, evolution of the Antarctic marine biota has been driven by the development of extreme cold temperatures. Because they live at very low and stable temperatures, Antarctic fishes of the suborder Nototheniodei are particularly attractive as models for understanding the mechanisms of biomolecular cold adaptation, or the compensatory restructuring of biochemical and physiological systems to preserve biological function in cold thermal regimes. Two interrelated and potentially co-evolved systems, the tubulins that form microtubules and the chaperonin-containing TCP1 (t-complex protein-1) complex (CCT) that assists the folding of tubulins, provide an unparalleled opportunity to elucidate these mechanisms. This research will yield new and important knowledge regarding: 1) cold adaptation of microtubule assembly and of chaperonin function; and 2) the co-evolutionary origin of tubulin-binding specificity by CCT. The first objective of this proposal is to determine the contributions of five novel amino acid substitutions found in Antarctic fish beta-tubulins to microtubule assembly at cold temperature. The second objective is to establish a chaperonin folding system in vitro using CCT purified from testis tissue of Antarctic fishes and to evaluate its thermal properties and mechanism. The third objective is to evaluate, through phylogenetically controlled contrasts, the hypothesis that CCT and its tubulin substrates from Antarctic fishes have co-evolved to function at cold temperatures. The broader impacts of this proposal include introduction of graduate and REU undergraduate students of Northeastern University to state-of-the-art biochemical, cellular, and molecular-biological research relevant to ecological and environmental issues of the Antarctic marine ecosystem. Because much of the research on the biogenesis and function of cold-adapted proteins will be performed in the field at Palmer Station, these students will gain invaluable experience in the practical considerations of expeditionary biological science. The research also will increase knowledge about molecular cold adaptation in one of the Earth's extreme environments, and hence is relevant to the formulation of refined hypotheses regarding potential extraterrestrial life on Mars or Europa. The cold-functioning chaperonin protein folding system will be of great value to the biopharmaceutical and biotechnological industries for use in folding insoluble proteins.
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