Reactivation of the male gametogenic expression program is tightly associated with the most malignant and metastasis-prone tumors and the emergence of aggressive sub-clones of tumor cells, which are highly resistant to stress-induced apoptosis. While the cancer-testis antigens (CTAs) CABYR and AKAP3/4 roles during gametogenesis and their importance for flagellar movement have gradually emerged, their function in cancer cells has remained obscure. In this study, we combine immunoprecipitation (IP), mass spectrometry (MS), and Western blot (WB) analysis to unravel their functional roles in therapy-resistant lung and ovary adenocarcinoma cells by identifying their interaction partners. CABYR variants were shown to oligomerize and interact with AKAP proteins to generate an HMW signal scaffold structure, which was found to bind several glycolytic enzymes and signal transducers. Forward and reverse IP experiments followed by WB confirmed interactions between CABYR and LDH, ALDO, PFK, TPI-1, GAPDH, ENO-1, and GSK3b. The transition from normoxic to hypoxic growth conditions disrupted the associations between glycolytic enzymes and the CABYR-AKAP signaling scaffold in the cancer cells, leading to a 3.2-fold increase in their production and secretion of lactic acid. Hypoxic growth conditions resulted in increased acetylation of lysine residues in both CTAs and triggered deacetylation of lysines in LDH and aldolase.
