LATS1 Regulates Mixed-Lineage Kinase 3 (MLK3) Subcellular Localization and MLK3-Mediated Invasion in Ovarian Epithelial Cells

Mixed-lineage kinase 3 (MLK3) activates mammalian mitogen-activated protein kinase (MAPK) signaling pathways in response to cytokines and stress stimuli. MLK3 is important for proliferation, migration, and invasion of different types of human tumor cells. We observed that endogenous MLK3 was localized to both the cytoplasm and the nucleus in immortalized ovarian epithelial (T80) and ovarian cancer cells, and mutation of arginines 474 and 475 within a putative MLK3 nuclear localization sequence (NLS) resulted in exclusion of MLK3 from the nucleus. The large tumor suppressor (LATS) Ser/Thr kinase regulates cell proliferation, morphology, apoptosis, and mitotic exit in response to cell-cell contact. RNA interference (RNAi)-mediated knockdown of LATS1 increased nuclear, endogenous MLK3 in T80 cells. LATS1 phosphorylated MLK3 on Thr477, which is within the putative NLS, and LATS1 expression enhanced the association between MLK3 and the adapter protein 14-3-3ζ. Thr477 is essential for MLK3-14-3-3ζ association and MLK3 retention in the cytoplasm, and a T477A MLK3 mutant had predominantly nuclear localization and significantly increased invasiveness of SKOV3 ovarian cancer cells. This study identified a novel link between the MAPK and Hippo/LATS1 signaling pathways. Our results reveal LATS1 as a novel regulator of MLK3 that controls MLK3 nuclear/cytoplasmic localization and MLK3-dependent ovarian cancer cell invasion.