PROTEOMICS OF HYPOTHERMIC ADAPTATION REVEALS THAT RBM3 ENHANCES MITOCHONDRIAL METABOLISM AND MUSCLE STEM-CELL DIFFERENTIATION

Abstract

Adaptation to hypothermic stress is important for skeletal muscle cells, but a 17 comprehensive knowledge of molecular mediators is lacking. We show that adaptation to 18 mild hypothermia (320C) improves the ability of skeletal muscle myoblasts to differentiate 19 into myotubes in vitro. We performed proteomic analysis of mouse myoblasts exposed to 20 mild hypothermia for various time points and identified dynamic changes in 21 mitochondrial metabolism and proteostasis. This revealed that RBM3, an RNA-binding 22 protein, increases progressively with acute and chronic exposure to hypothermic stress, 23 and is necessary for the enhanced differentiation upon hypothermic adaptation. We also 24 demonstrate that overexpression of RBM3 at physiological temperatures is sufficient to (i) 25 enhance mitochondrial metabolism as judged by a decrease in the AMPK energy-sensing 26 pathway, (ii) increase levels of proteins associated with translation and increase levels of 27 4E-BP1 phosphorylation, (iii) increase stem cell markers (MyoD1, PAX7), and improve 28 differentiation of myoblasts from both young and aged mice.