Despite the central role of mitogen-activated protein kinase (MAPK) p38 in driving pathological inflammation - including during pulmonary injury - effective therapeutic targeting remains elusive. Atypical p38 signaling, mediated by interaction with the adaptor protein Tumor Growth Factor β Activated Kinase 1 (TAK1) Binding Protein 1 (TAB1) is activated only in pathological responses, representing a selective and alternative target during pulmonary inflammation. Our prior work established that G-protein coupled receptors (GPCRs) rapidly activate atypical p38 signaling in primary human endothelial cells, inducing vascular disruption and inflammatory cytokine production, potentially implicating atypical signaling in acute lung injury (ALI). However, there are key gaps in our knowledge about how atypical signaling differentiates from canonical activation and how this pathway impacts human health. Emerging evidence suggests that kinase activity is spatiotemporally regulated, and downstream signaling is mediated by spatially restricted substrate access. We hypothesize that differences in kinase localization and activity may be the driving factor behind the physiological effects of atypical p38 signaling. This work encapsulates the current knowledge on kinase localization dynamics and atypical signaling as emerging fields of study. Our studies branch into two projects, where we seek to elucidate the spatial dynamics of p38 activity and uncover the first known roles of atypical signaling in infectious pulmonary inflammatory disease progression. To accomplish this, we use a combination of in vitro studies utilizing Fluorescence Resonance Energy Transfer (FRET) microscopy paired with genetically encoded kinase activity reporters and in vivo studies of a mouse model of influenza infection. Our results indicate that canonical, MKK3/6-driven p38 activity via osmotic stress results in rapid, transient kinase signaling that is strongest in the nucleus. In contrast, atypical signaling by GPCR agonists initiates sustained signaling primarily in the cytosol and endosome. Additionally, we report that systemic knock-in mice deficient for TAB1-p38 interaction are protected from influenza-induced ALI as measured by weight loss curves, histopathological scoring, mRNA expression, and immune recruitment to the lung. The collective discoveries in this dissertation demonstrate the potential therapeutic benefit of targeting atypical p38 signaling and lay the groundwork for future studies exploring how spatially restricted kinase/substrate interaction contributes to pathological outcomes.