Objective: Long noncoding RNAs (lncRNAs) are emerging as important regulators of diverse biological processes. Recent work has demonstrated that the inducible lncRNA Blnc1 stimulates thermogenic gene expression during brown and beige adipocyte differentiation. However, whether Blnc1 is functionally conserved in humans has not been explored. In addition, the molecular basis of the Blnc1 ribonucleoprotein complex in thermogenic gene induction remains incompletely understood. The aims of the current study were to: i) investigate functional conservation of Blnc1 in mice and humans and ii) elucidate the molecular mechanisms by which Blnc1 controls the thermogenic gene program in brown adipocytes.
Methods: Full-length human Blnc1 was cloned and examined for its ability to stimulate brown adipocyte differentiation. Different truncation mutants of Blnc1 were generated to identify functional RNA domains responsible for thermogenic gene induction. RNA-protein interaction studies were performed to delineate the molecular features of the Blnc1 ribonucleoprotein complex.
Results: Blnc1 is highly conserved in mice and humans at the sequence and function levels, both capable of stimulating brown adipocyte gene expression. A conserved RNA domain was identified to be required and sufficient for the biological activity of Blnc1. We identified hnRNPU as an RNA-binding protein that facilitates the assembly and augments the transcriptional function of the Blnc1/EBF2 ribonucleoprotein complex.
Conclusions: Blnc1 is a conserved lncRNA that promotes thermogenic gene expression in brown adipocytes through formation of the Blnc1/hnRNPU/EBF2 ribonucleoprotein complex.
Keywords: ATP5A, ATP synthase, H+ transporting, mitochondrial F1 complex, alpha 1; BAT, brown adipose tissue; Blnc1; Brown adipocyte differentiation; Brown fat; Cox7a1, cytochrome c oxidase subunit 7A1; Dio2, deiodinase, iodothyronine type II; EBF2; EBF2, early B cell factor 2; Elovl3, elongation of very long chain fatty acids like 3; FABP4, fatty acid binding protein 4; PPARγ, peroxisome proliferator-activated receptor gamma; Ppargc1a, peroxisome proliferator-activated receptor gamma coactivator 1-alpha; Pparα, peroxisome proliferator-activated receptor alpha; Prdm16, PR domain zinc finger protein 16; RACE, rapid amplification of cDNA ends; SDHB, succinate dehydrogenase complex iron sulfur subunit B; Thermogenesis; UQCRC2, ubiquinol-cytochrome c reductase core protein II; Ucp1, uncoupling protein 1; lncRNA.