Identifying protein domain(s) and interactors necessary for Mcrip2 function in brown adipocytes

High in mitochondria and oxidative capacity, brown adipose tissue (BAT) produces heat through the process of adaptive thermogenesis. Understanding the mechanisms that enable the high oxidative metabolic capacity in BAT can provide insight into and enhance current therapeutic approaches targeting conditions of metabolic dysfunction. Recently, the Kralli lab has identified a gene implicated in BAT thermogenesis that encodes the “mitogen-activated protein kinase (MAPK)-regulated corepressor interacting protein 2” (Mcrip2). Preliminary studies show Mcrip2 regulates the mRNA levels of genes involved in oxidative metabolism, including those involved in oxidative phosphorylation ( Uqcrfs1), fatty acid oxidation ( Acat1, Cpt2), and branched-chain amino acid catabolism ( Bcat2, Bckdha, Ivd, Echs1, Pcca). Interestingly, unbiased proteomic experiments in HEK203 cells have identified multiple Mcrip2 interactors (including Ddx6, Atxn2, Atxn2l) involved in RNA processing, translation and decay, suggesting that this regulation could be post-transcriptional. However, how Mcrip2 post-transcriptionally regulates these genes and the identity of the interactors Mcrip2 constitutes its effects through remains unknown. To elucidate the mechanism of action of Mcrip2, the lab created eight C-terminal HA-tagged Mcrip2 mutant constructs, with mutations targeting different evolutionarily conserved regions of the protein. These mutants will be expressed in primary brown adipocytes lacking Mcrip2 to determine if they complement wild-type and will also be used for co-immunoprecipitation. Results from such studies may provide further insight into the role of Mcrip2 in adipocyte oxidative metabolism and, more broadly, in BAT function. NIH. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.