Profiling mouse brown and white adipocytes to identify metabolically relevant small ORFs and functional microproteins
Thomas F. Martínez, Sally Lyons‐Abbott, Angie L. Bookout, Eduardo Vieira de Souza, Cynthia J. Donaldson, Joan Vaughan, Calvin J. Lau, Ariel Abramov, Arian F. Baquero, Karalee Baquero, Dave Friedrich, Justin Huard, Ray Davis, Bong-Chul Kim, Ty Koch, Aaron J. Mercer, Ayesha Misquith, Sara A. Murray, Sakara Perry, Lindsay K. Pino, Christina A. Sanford, Alex Simon, Yu Zhang, Garrett Zipp, Cristiano Valim Bizarro, Maxim N. Shokhirev, Andrew J. Whittle, Brian C. Searle, Michael J. MacCoss, Alan Saghatelian, Christopher A. Barnes
Abstract
Microproteins (MPs) are a potentially rich source of uncharacterized metabolic regulators. Here, we use ribosome profiling (Ribo-seq) to curate 3,877 unannotated MP-encoding small ORFs (smORFs) in primary brown, white, and beige mouse adipocytes. Of these, we validated 85 MPs by proteomics, including 33 circulating MPs in mouse plasma. Analyses of MP-encoding mRNAs under different physiological conditions (high-fat diet) revealed that numerous MPs are regulated in adipose tissue in vivo and are co-expressed with established metabolic genes. Furthermore, Ribo-seq provided evidence for the translation of Gm8773, which encodes a secreted MP that is homologous to human and chicken FAM237B. Gm8773 is highly expressed in the arcuate nucleus of the hypothalamus, and intracerebroventricular administration of recombinant mFAM237B showed orexigenic activity in obese mice. Together, these data highlight the value of this adipocyte MP database in identifying MPs with roles in fundamental metabolic and physiological processes such as feeding.