Winter cold hardiness of insects typically involves one of two major strategies for survival below 0°C: freeze avoidance and freeze tolerance. The two strategies have some common features, including the accumulation of high concentrations of cryoprotectant polyols and the frequent occurrence of diapause. Entry into the hypometabolic state of diapause requires coordinated suppression of major ATP-consuming metabolic processes, and ion motive ATPases are important targets for regulation. This study documents the suppression of sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) activity in the overwintering larvae of two cold-hardy species, the freeze-avoiding gall moth Epiblema scudderiana and the freeze-tolerant gall fly Eurosta solidaginis. Activity was reduced despite a lack of change in SERCA protein levels in E. solidaginis larvae over the winter and a six- to eightfold increase in SERCA protein in E. scudderiana. This implicated posttranslational modification as the mechanism of SERCA suppression, and in vitro incubations indicated that enzyme phosphorylation by protein kinases A, G, or C strongly reduced enzyme activity. A stable reduction in SERCA activity was also seen in cold-acclimated larvae of both species compared with 15°C controls, with significant changes in the kinetic parameters of the E. scudderiana enzyme (e.g., Km ATP was 3.2-fold higher in - 20°C-acclimated larvae) that were consistent with reduced enzyme function at low temperature. Epiblema scudderiana SERCA was also subject to regulation by differential temperature effects (Arrhenius activation energy increased by approximately threefold below 10°C) and by seasonal changes in the levels of a SERCA inhibitor protein, phospholamban.
Physiological and Biochemical Zoology
Department of Chemistry

McMullen, D.C. (David C.), Ramnanan, C.J. (Christopher J.), & Storey, K. (2010). In cold-hardy insects, seasonal, temperature, and reversible phosphorylation controls regulate sarco/endoplasmic reticulum Ca 2+-ATPase (SERCA). Physiological and Biochemical Zoology, 83(4), 677–686. doi:10.1086/653489