Adipose tissue (AT) accumulation, particularly intra-abdominally, is tightly associated with obesity-related co-morbidities. Since AT is increasingly-appreciated to regulate whole-body metabolism, this connection has been attributed to obesity-triggered AT stresses, that in-turn cause adipose dysfunction. We reasoned that the "translation" of various AT stresses into dysfunction involves activation of MAPkinases in human intra-abdominal fat. Testing mRNA, phopho- and total protein levels and comparing omental and subcutaneous fat from obese and non-obese persons, we propose a functional MAPkinase signaling cascade involving the MAP3K ASK1(MAP3K5), but not Tak1 or MLK3, the MAP2K's MKK4,3,6 (but not MKK7), and the stress-activated JNK and p38MAPK. Beyond the well-studied phosphorylation-mediated cascade, the expression of ASK1 was up-regulated in omental fat, particularly in persons with intra-abdominal fat distribution. Importantly, multivariate model demonstrated omental ASK1mRNA as an independent predictor of whole-body insulin resistance. In cultured adipocytes ASK1 mRNA increased in response to inflammation and oxidative stress (but not inducers of ER stress), a response fully inhibitable by actinomycinD. We therefore set to explore the transcriptional regulation of ASK1, following its putative regulation by E2F transcription factors. Omental E2F1 mRNA and protein were increased in obesity. MEFs-derived adipocyte-like cells from E2F1-KO mice showed decreased ASK1 expression and activation compared to WT-MEFs. Furthermore, the responsiveness of the human-ASK1 promoter required the co-overexpression of E2F1, and mutating the putative E2F1 binding site in the promoter decreased promoter activation. Yet, fully preventing ASK1 promoter required the addition of JNK inhibitor. Finally, ChIP studies demonstrated binding of E2F1 to the ASK1 promoter in human fat, correlating with obesity. In summary, a combined phosphorylation and E2F1-based transcriptional regulation of an ASK1-based pathway operates to sensitize intra-abdominal fat to obesity-related stresses.