Edited by Linda SpremulliAlthough exercise is linked with improved health, the specific molecular mechanisms underlying its various benefits require further clarification. Here we report that exercise increases the nuclear localization and activity of p53 by acutely down-regulating coiled-coil-helix-coiled-coil-helix domain 4 (CHCHD4), a carrier protein that mediates p53 import into the mitochondria. This response to exercise is lost in transgenic mice with constitutive expression of CHCHD4. Mechanistically, exerciseinduced nuclear transcription factor FOXO3 binds to the CHCHD4 promoter and represses its expression, preventing the translocation of p53 to the mitochondria and thereby increasing p53 nuclear localization. The synergistic increase in nuclear p53 and FOXO3 by exercise can facilitate their known interaction in transactivating Sirtuin 1 (SIRT1), a NAD ؉ -dependent histone deacetylase that mediates adaptation to various stresses. Thus, our results reveal one mechanism by which exercise could be involved in preventing cancer and potentially other diseases associated with aging.The nuclear transcriptional activities of p53, well established to play a critical role in tumor suppression, are dynamically regulated by multiple factors, including posttranslational modifications and protein-protein interactions (1-3). Accumulating evidence also shows that p53 localization in non-nuclear subcellular compartments, such as the cytosol and mitochondria, can regulate cellular processes like autophagy and metabolism (4 -6). Although stress-mediated p53 translocation to the mitochondria and its role in cell death have been well delineated, the effect of p53 translocation into mitochondria under normal states is less clear (7,8). Thus, understanding the regulation of p53 partitioning into the mitochondria under physiologic conditions may provide new insights into its tumor-suppressive functions.We reported previously that CHCHD4, 2 the mammalian homolog of the yeast mitochondrial disulfide relay system carrier Mia40, interacts with p53 and mediates its translocation into mitochondria (9). Unlike other mechanisms by which proteins can be imported into mitochondria, the disulfide relay system is coupled to respiration, perhaps reflecting a homeostatic mechanism by which the import of p53 can be regulated by mitochondrial activity (10, 11). Subsequently, p53 import into mitochondria by CHCHD4 facilitates the repair of oxidative mtDNA damage, complementing the nuclear role of p53 in regulating mitochondrial respiration (9, 12, 13). However, these observations also raise questions regarding the physiological significance of the partitioning of p53 between the nucleus and mitochondria and whether such a mechanism can be demonstrated in vivo under normal conditions.Exercise elicits many complex physiological responses, including the induction of specific transcription factors such as PGC-1␣ and peroxisome proliferator-activated receptors, which transactivate mitochondrial biogenesis genes (14, 15). As a mitochondrial protein ...