Surfactant proteins maintain lung function through their actions to reduce alveolar surface tension and control of innate immune responses in the lung. The ubiquitin proteasome pathway is responsible for the degradation of majority of intracellular proteins in eukaryotic cells, and proteasome dysfunction has been linked to the development of neurodegenerative, cardiac and other diseases. Proteasome function is impaired in interstitial lung diseases associated with SP-C mutation mapping to the BRICHOS domain located in the proSP-C protein. In this study we determined the effects of proteasome inhibition on surfactant protein expression in H441 and MLE-12 lung epithelial cells to understand the relationship between proteasome dysfunction and surfactant protein gene expression. Proteasome inhibitors, lactacystin and MG132, reduced the levels of SP-A, SP-B and SP-C mRNAs in a concentration-dependent manner in H441 and MLE-12 cells. In H441 cells, lactacystin and MG132 inhibition of SP-B mRNA was associated with similar decreases in SP-B protein and the inhibition was due to inhibition of gene transcription. Proteasome inhibitors decreased TTF-1/Nkx2.1 DNA binding activity and the reduced TTF-1 DNA binding activity was due to reduced expression levels of TTF-1 protein. These data indicate that the ubiquitin proteasome pathway is essential for the maintenance of surfactant protein gene expression and disruption of this pathway inhibits surfactant protein gene expression via reduced expression of TTF-1 protein.