Analysis of changes in mRNA translation during adipogenesis and its regulation by mTORC1

Abstract

Understanding the biology of adipose tissue and the molecular mechanisms involved in adipogenesis is essential for the development of therapies aiming to control obesity and associated diseases. Despite many studies addressing transcription regulation, we still do not know much about the control of mRNA translation into proteins. The development of ribosome profiling technique that enables the analysis of ribosome positions and indicates which mRNAs are actually being translated into proteins has brought new interest in this area, specially due to the use of translation initiation inhibitors that allow the identification of alternative translation initiation. In this proposal, we aim to characterize the changes in the rates of mRNA translation into proteins during preadipocyte differentiation into mature adipocytes that are independent of changes in transcription. It is also our major goal to evaluate the role of mTOR in the regulation of mRNA translation during adipogenesis. For this we will carry a deep analysis of mRNA transcription and translation in different time points over 3T3-L1 differentiation into mature adipocytes by using a combination of global translation initiation sequencing and RNA-seq. Accordingly, 3T3-L1 cells will be harvested at 0 h, 4 h, 12 h, 24 h and 48 h after the addition of the differentiation cocktail (insulin, IBMX, dexamethasone) in the absence or presence of mTOR inhibitor torin for global translation initiation sequencing and RNA-seq. (AU)