Physiological adaptation through acclimation is one way to cope with temperature changes. Biochemical studies on acclimation responses in ectotherms have so far mainly investigated consequences of short-term acclimation at the adult stage and focussed on adaptive responses. Here we assessed the consequences of developmental and adult rearing at low (12°C), benign (25°C) and high (31°C) temperatures in Drosophila melanogaster. We assessed cold and heat tolerance and obtained detailed proteomic profiles of flies from the three temperatures. The proteomic profiles provided a holistic understanding of the underlying biology associated with both adaptive and non-adaptive temperature responses. Results show strong benefits and costs across tolerances: rearing at low temperature increased adult cold tolerance and decreased adult heat tolerance and vice versa with development at high temperatures. In the proteomic analysis we were able to identify and quantify a large number of proteins compared to previous studies on ectotherms (1440 proteins across all replicates and rearing regimes), enabling us to extend the proteomic approach using enrichment analyses. This gave us both detailed information on individual proteins as well as pathways affected by rearing temperature, pinpointing mechanisms likely responsible for the strong costs and benefits of rearing temperature on functional phenotypes. Several well-known heat shock proteins as well as proteins not previously associated with thermal stress were among the differentially expressed proteins. Upregulation of proteasome proteins was found to be an important adaptive process at high stressful rearing temperatures, and occurs at the expense of downregulation of basal metabolic functions.