Respiratory syncytial virus (RSV) is a significant human pathogen, causing acute lower respiratory illness in millions of infants, and exacerbating asthma, bronchiolitis and pneumonia in the elderly and immunosuppressed adults. Although the clinical manifestations are known, the pathogenesis of RSV infection is not well understood. The involvement of host cell mitochondria, as part of anti-viral immune responses, remained unclear. With recent proteomics indicating RSV’s impact on many nuclear-encoded mitochondrial proteins, we are evaluating for the first time the association between RSV and mitochondria in A549 adenocarcinomic human alveolar basal epithelial cells. Our studies focused first on documenting RSV-induced changes in mitochondrial morphology and respiratory function over the first 48 hours of infection, across different multiplicities of infection (MOIs). Mechanistic insights are obtained using different mutant forms of RSV and siRNA strategies to probe which viral encoded proteins contribute to altered mitochondria. Our results will pave the way for possible therapeutic interventions and anti-RSV strategies in the future.