Background: We have previously identified mitogen activated protein kinase activated protein kinase 2 (MK2) is required for caspase-3 nuclear translocation in the execution of apoptosis, however, little is known of the underlying mechanisms. Therefore, we sought to determine the role of kinase and non-kinase functions of MK2 in promoting nuclear translocation of caspase-3. Methods: We identified two non-small cell lung cancer cell lines for use in these experiments based on low MK2 expression. Wild-type, enzymatic and cellular localization mutant MK2 constructs were expressed using adenoviral infection. Cell death was evaluated by flow cytometry. In addition, cell lysates were harvested for protein analyses. Phosphorylation of caspase-3 was determined using two-dimensional gel electrophoresis followed by immuno-blotting and in vitro kinase assay. Association between MK2 and caspase-3 was evaluated using proximity-based biotin ligation assays and co-immunoprecipitation. Results: Overexpression of MK2 resulted in nuclear translocation of caspase-3 and caspase-3 mediated apoptosis. MK2 directly phosphorylates caspase-3, however, phosphorylation status of caspase-3 or MK2-dependent phosphorylation of caspase-3 did not alter caspase-3 activity. The enzymatic function of MK2 was dispensable in nuclear translocation of caspase-3. MK2 and caspase-3 associated together and a non-enzymatic function of MK2, chaperoned nuclear trafficking, is required for caspase-3 mediated apoptosis. Conclusion: Taken together, our results demonstrate a non-enzymatic role for MK2 in the nuclear translocation of caspase-3. Furthermore, MK2 may function as a molecular switch in regulating the transition between the cytosolic and nuclear functions of caspase-3.

Rationale: Non-small cell lung cancers (NSCLC) demonstrate intrinsic resistance to cell death, even after chemotherapy. Previous work suggested defective nuclear translocation of active caspase-3 in observed resistance to cell death. We have identified mitogen activated protein kinase activated protein kinase 2(MK2) is required for caspase-3 nuclear translocation in the execution of apoptosis in endothelial cells. Objective: To determine MK2 expression in NSCLCs and the association between MK2 and clinical outcomes in NSCLC patients. Methods: Clinical and MK2 mRNA data were extracted from two demographically distinct NSCLC clinical cohorts, North American (The Cancer Genome Atlas, TCGA) and East Asian (EA). Tumor responses following first round of chemotherapy were dichotomized as Clinical Response (complete response, partial response and stable disease) or Progression of Disease. Multivariable survival analyses were performed using Cox proportional hazard ratios, logistic regression odd ratios and Kaplan-Meier curves. Results: NSCLC exhibited lower MK2 expression than SCLC cell lines. In patients, lower tumor MK2 transcript levels were observed in those presenting with late-stage NSCLC. Higher MK2 expression was associated with clinical response following initial chemotherapy and independently associated with improved two-year survival in two distinct cohorts, 0.52(0.28-0.98) and 0.1(0.01-0.81), TCGA and EA respectively, even after adjusting for common oncogenic driver mutations. Survival benefit of higher MK2 expression was unique to lung adenocarcinoma when comparing across cancers. Conclusion: This study implicates MK2 in apoptosis resistance in NSCLC and suggests prognostic value of MK2 transcript levels in patients with lung adenocarcinoma.