Naked mole rats are among the most hypoxia‐tolerant mammals identified. This tolerance is partially achieved by their ability to coordinately reduce metabolic and ventilatory rates by > 85% in acute hypoxia. Our understanding of the physiological mechanisms that enable their hypoxia‐tolerance is improving; however, the cellular mechanisms that mediate these responses remain poorly understood. Hypoxia‐inducible factor‐1α (HIF‐1α) is a transcription factor that is central to hypoxic signaling at the cellular level. In most mammals, HIF‐1α is degraded in normoxia but is stabilized when oxygen levels are reduced. Stabilization of HIF‐1α leads to upregulation of a multitude of genes that promote survival in hypoxia. A recent analysis of the naked mole rat genome revealed several mutations to HIF‐1α. These mutations are predicted to prevent HIF‐1α degradation during normoxia, which would chronically upregulate HIF‐1α‐mediated signaling pathways. We hypothesized that endogenous activation of HIF‐1α‐mediated signaling contributes to the remarkable hypoxia‐tolerance of naked mole rats by modulating their metabolic rate. To test our hypothesis, we used pharmacological tools to manipulate HIF‐1α expression in naked mole rats and mice and measured metabolic and ventilatory responses to an acute hypoxic challenge (7% O2 for 1hr) in awake and unrestrained subjects using plethysmography and respirometry. Pharmacological manipulation of HIF‐1α expression was confirmed by measuring changes in the expression of several downstream gene targets of HIF‐1α. In naked mole rats, blocking HIF‐1α had only minor effects on ventilation but drastically blunted the hypoxic metabolic response. As a result, the naked mole rat air convection requirement (ACR) increased 2‐fold in normoxia and 3‐fold in hypoxia, indicating that naked mole rats were hyperventilating and thus less tolerant of hypoxic stress. Conversely, activating HIF‐1α in mice reduced ventilation and metabolic rate in both normoxia and hypoxia and also reduced the hypoxic increase in the murine ACR, indicating that HIF‐1α‐agonist‐treated mice were more tolerant of hypoxic stress. We conclude that endogenous upregulation of HIF‐1α signaling contributes to the hypoxia‐tolerance of naked mole rats, and we demonstrate that this mechanism can be translated to hypoxia‐intolerant mice to induce a hypoxia‐tolerant phenotype.Support or Funding InformationNatural Sciences and Engineering Research Council of Canada, Canada Research Chairs Program, Parker B Francis Foundation