Abstract Adenoid Cystic Carcinoma (ACC), the second most common malignancy of salivary glands, is a rare tumor with bleak prognosis for which therapeutic targets are unavailable. Recently, we used RNA-sequencing (RNA-Seq) to analyze low-quality RNA from archival, formaldehyde-fixed, paraffin-embedded samples (1). In addition to detecting the most common ACC translocation, t(6;9) fusing the MYB proto-oncogene to NFIB, we also detected previously unknown t(8;9) and t(8;14) translocations fusing the MYBL1 gene to the NFIB and RAD51B genes, respectively. Mutated versions of c-Myb are potent oncoproteins, and Myb proteins are over- or highly-expressed in a wide variety of human tumors. Interestingly, ACC tumors with MYB and MYBL1 translocations displayed similar gene expression profiles, and the combined MYB and MYBL1 expression correlated with outcome, suggesting that the related Myb proteins are interchangeable oncogenic drivers in ACC. While RNA-Seq provides information about gene fusions, alternative RNA splicing and gene expression signatures, this type of data does not address how translocations alter transcription factor function. Previous studies in our lab, and others, have demonstrated that Myb proteins contain C-terminal regulatory domains and are known to interact with several proteins via C-terminal motifs (2). These regulatory and interaction sites are lost upon chromosomal translocation in ACC; however, the downstream affect of this loss is not understood. Do the translocations merely relieve Myb transcription factors of repressive regulatory signals, thereby globally increasing Myb-dependent gene expression or, does the altered protein allow novel protein-protein interactions which re-target Myb transcription factors? Furthermore, the most frequent translocations involve the relocation of a large enhancer regions. What role does the recruitment of novel enhancer regions play in up-regulating Myb protein expression? Using a combination of ChIP-Seq and digital genomic footprinting, we are attempting to understand the mechanism of regulatory rewiring observed in ACC tumors.