A LATS2 and ALKBH5 positive feedback loop supports their oncogenic roles.

N(6)-methyladenosine (m⁶A) critically regulates RNA dynamics in various biological processes. The m⁶A demethylase ALKBH5 promotes tumorigenesis of glioblastoma, while the intricate web that orchestrates its regulation remains enigmatic. Here, we discover that cell density affects ALKBH5 subcellular localization and m⁶A dynamics. Mechanistically, ALKBH5 is phosphorylated by the large tumor suppressor kinase 2 (LATS2), preventing its nuclear export and enhancing protein stability. Furthermore, phosphorylated ALKBH5 reciprocally erases m⁶A from LATS2 mRNA, thereby stabilizing this transcript. Unexpectedly, LATS2 depletion suppresses glioblastoma stem cell self-renewal independent of yes-associated protein activation. Additionally, deficiency in either LATS2 or ALKBH5 phosphorylation impedes tumor progression in mouse xenograft models. Moreover, high levels of LATS2 expression and ALKBH5 phosphorylation are associated with tumor malignancy in patients with gliomas. Collectively, our study unveils an oncogenic positive feedback loop between LATS2 and ALKBH5, revealing a non-canonical branch of the Hippo pathway for RNA processing and suggesting potential anti-cancer interventions. [Display omitted] • LATS2 phosphorylates ALKBH5 at Ser361, ensuring its nuclear localization and stability • Phosphorylated ALKBH5 in the nucleus stabilizes LATS2 mRNA by demethylating m⁶A • The LATS2/ALKBH5 positive feedback loop is crucial for their oncogenic roles Cao et al. show that the MST1/2-LATS2 axis is active in glioblastoma, phosphorylates ALKBH5, and safeguards its nuclear localization and protein stability. In turn, ALKBH5 stabilizes LATS2 mRNA by demethylating m⁶A, thereby forming a positive feedback loop to promote glioblastoma progression. [ABSTRACT FROM AUTHOR]