Coordinated regulation of Barrier-to-Autointegration Factor (BAF) phosphorylation by protein phosphatases PP2A and PP4

Phosphorylation is crucial for the regulation of Barrier-to-Autointegration Factor (BAF), a conserved nuclear envelope (NE)-associated protein that, among its diverse functions, binds chromatin and mediates its anchoring to the NE. In interphase, cycles of BAF phosphorylation, which inhibits chromatin binding, and dephosphorylation, which restores it, are essential for the dynamic anchoring of chromatin to the NE. During mitosis, BAF becomes highly phosphorylated and dissociates from chromatin, except for a small centromeric pool (cenBAF). Upon mitotic exit, BAF is dephosphorylated and re-associates with chromosomes. Two Ser/Thr protein phosphatases, PP2A and PP4, are known to dephosphorylate BAF, but their specific roles in regulating BAF phosphorylation are not yet fully understood. Here, we show that PP2A is the main phosphatase driving BAF dephosphorylation in interphase, whereas PP4 plays a minor role. During mitosis, both PP2A and PP4 localize to centromeres; however, only PP4 directly interacts with cenBAF, suggesting that it maintains this fraction in an unphosphorylated state. Notably, PP4s centromeric localization depends on PP2A. At mitotic exit, PP2A dephosphorylates the large pool of mitotically phosphorylated BAF, while PP4 controls the timing of its re-association with chromatin. Together, these findings reveal a coordinated interplay between PP2A and PP4 in controlling BAF dephosphorylation and function.