GNL3 regulates replication origin firing and protects stalled replication forks

Abstract

DNA replication requires specific proteins that protect replication forks and so prevent the formation of DNA lesions that may damage the genome. Identification of new proteins involved in these processes is essential to understand how cancer cells tolerate DNA lesions. Here we show that human GNL3/nucleostemin, a GTP-binding protein localized mostly in the nucleolus and highly expressed in cancer cells, prevents nuclease-dependent resection of nascent DNA in response to exogenous replication stress. We demonstrate that inhibition of origin firing decreases this resection, indicating that the increased replication origin firing seen upon GNL3 depletion mainly accounts for the observed DNA resection. We show that GNL3 and DNA replication initiation factor ORC2 interact in the nucleolus and that the concentration of GNL3 in the nucleolus is required to limit DNA resection in response to replicative stress. We propose that the accurate control of origin firing by GNL3, possibly through the regulation of ORC2 sub-nuclear localization, is critical to prevent nascent DNA resection in response to replication stress.