LAUSR

Abstract Previous studies have shown that SIRT2 plays a role in mitosis through deacetylating specific downstream targets. However, the upstream regulation of SIRT2 activity has been relatively unexplored. In this study, we provide evidence that NAD(P)H:quinone oxidoreductase 1 (NQO1) interacts with and activates SIRT2 in an NAD‐dependent manner. Strong protein‐protein interaction and co‐localization of the two proteins during mitosis is required to maintain an active NQO1‐SIRT2 axis which is critical for successful completion of mitosis. This is evident by the observed delay in mitotic exit in cells upon NQO1 inhibition. Mechanistically, this phenotype can be explained by the decrease in APC/C complex activity resulting from decreased SIRT2 deacetylation activity. Furthermore, we show that this newly established role of NQO1 has an impact on how cancer cells may respond to mitotic stress. In this regard, both pharmacologic and genetic NQO1 inhibition increases sensitivity to anti‐mitotic drugs functioning as microtubule poisons by inducing mitotic arrest and allowing cells to accumulate cell death signals. Therefore, the significant prognostic value of NQO1 in predicting outcome of cancer patients might be explained in part due to the functional contribution of NQO1‐SIRT2 axis to mitotic stress. Altogether, this novel mechanism of action further supports the pleiotropic biological effects exerted by NQO1 in addition to its antioxidant function and it might provide the basis for expanding the therapeutic potential of NQO1 inhibition towards increasing sensitivity to standard treatments. Graphical abstract Figure. No caption available. HighlightsNQO1 is an upstream positive regulator of SIRT2 activity by providing NAD+.NQO1 colocalizes with SIRT2 during mitosis and regulates mitotic progression.SIRT2‐directed acetylation status of APC/C complex provides the mechanistic link.The NQO1/SIRT2 axis affects response of cancer cells to anti‐mitotic compounds/drugs.