16Cold stenothermal insects living in glacier-fed streams are stressed by temperature variations 17 resulting from glacial retreat during global warming. The molecular aspects of insect response to 18 environmental stresses remain largely unexplored. The aim of this study was to expand our 19 knowledge of how a cold stenothermal organism controls gene expression at the transcriptional, 20 translational, and protein level under warming conditions. Using the chironomid Diamesa tonsa as 21 target species and a combination of RACE, qPCR, polysomal profiling, western blotting, and 22 bioinformatics techniques, we discovered a new molecular pathway leading to previously overlooked 23 adaptive strategies to stress. We obtained and characterized the complete cDNA sequences of three 24 heat shock inducible 70 (hsp70) and two members of heat-shock cognate 70 (hsc70). Strikingly, we 25 showed that a novel pseudo-hsp70 gene encoding a putative long noncoding RNA (lncRNA) which 2 26 is transcribed during thermal stress, acting as a ribosome sponge to provide post-transcriptional 27 control of HSP70 protein levels. The expression of the pseudo-hsp70 gene and its function suggest 28 the existence of a new and unexpected mechanism to cope with thermal stress: lowering the pace of 29 protein production to save energy and optimize resources for recovery. 30 31 change 3 33 4 58machinery of the cell [15, 16, 17]. Also, under non-stressful conditions HSPs facilitate the correct 59 folding of proteins during translation and their transport across membranes [18, 19]. Among HSPs 60 the 70 kDa family, consisting of inducible (HSP70) and constitutive (heat shock cognate, HSC70) 61 forms, is the most studied in relation to thermal stress and has been found in all organisms 62 investigated to date [20, 21]. [22] and [13] demonstrated that HSC70 plays a role in cold resistance 63 for D. cinerella gr. larvae and that the HSR is comprised of a strong transcriptional boost of the 64 inducible HSP70 gene at a temperature six times higher than that at which they live in nature. The 65 involvement of HSC70 and HSP70 in cold and heat tolerance was observed in other cold adapted 66 chironomids such as adults of Belgica antarctica [23] and larvae of Pseudodiamesa branickii [24]. 67 Knowledge of how these insects control gene expression at the transcriptional, translational and 68 protein level under heating is still under-explored. The present study aims to address this using a 69 multi-level approach to study HSR at the transcriptional, translational and protein level in D. tonsa. 70 This is particularly important because studying changes in gene expression only at the transcriptional 71 level may be misleading [25, 26] given the poor average correlation between protein and transcript 72 [27, 28] due to post-transcriptional controls of gene expression. Here we hypothesize that, similar to 73 observations in higher eukaryotes, post-transcriptional control of hsp70 and hsc70 gene expression 74 in insects may exist. In line with this hypothesis, we i...