An Aurora B-RPA signaling axis secures chromosome segregation fidelity

Abstract

AbstractErrors in chromosome segregation underlie genomic instability associated with cancers. Resolution of replication and recombination intermediates and protection of vulnerable single-stranded DNA (ssDNA) intermediates during mitotic progression requires the ssDNA binding protein Replication Protein A (RPA). However, the mechanisms that regulate RPA specifically during unperturbed mitotic progression are poorly resolved. RPA is a heterotrimer composed of RPA70, RPA32 and RPA14 subunits and is predominantly regulated through hyperphosphorylation of RPA32 in response to DNA damage. Here, we have uncovered a mitosis-specific regulation of RPA by Aurora B kinase. Aurora B phosphorylates Ser-384 in the DNA binding domain B of the large RPA70 subunit and highlights a mode of regulation distinct from RPA32. Disruption of phosphorylation markedly affects cell viability and causes defects in chromosome segregation with hypersensitivity to DNA damaging agents. Phosphorylation at Ser-384 remodels the protein interaction domains of RPA and facilitates formation of higher density filaments. Furthermore, phosphorylation impairs RPA binding to DSS1 that likely suppresses homologous recombination during mitosis by preventing recruitment of DSS1-BRCA2 to exposed ssDNA. We showcase a critical RPA-Aurora B signaling axis in mitosis that is essential for maintaining genomic integrity.