Although human nucleoporin Tpr is frequently deregulated in cancer, its roles are poorly understood. Here we show that Tpr depletion generates transcription-dependent replication stress, DNA breaks, and genomic instability. DNA fiber assays and electron microscopy visualization of replication intermediates show that Tpr deficient cells exhibit slow and asymmetric replication forks under replication stress. Tpr deficiency evokes enhanced levels of DNA-RNA hybrids. Additionally, complementary proteomic strategies identify a network of Tpr-interacting proteins mediating RNA processing, such as MATR3 and SUGP2, and functional experiments confirm that their depletion trigger cellular phenotypes shared with Tpr deficiency. Mechanistic studies reveal the interplay of Tpr with GANP, a component of the TREX-2 complex. The Tpr-GANP interaction is supported by their shared protein level alterations in a cohort of ovarian carcinomas. Our results reveal links between nucleoporins, DNA transcription and replication, and the existence of a network physically connecting replication forks with transcription, splicing, and mRNA export machinery.

Centro Andaluz de Biología Molecular y Medicina Regenerativa CABIMER Universidad de Sevilla CSIC Seville Spain

Danish Cancer Society Research Center Copenhagen Denmark

Division of Genome Biology Department of Medical Biochemistry and Biophysics Science for Life Laboratory Karolinska Institute Stockholm Sweden

Genome and Cell Integrity Laboratory CSIR Indian Institute of Toxicology Research Lucknow India

IFOM Fondazione Istituto FIRC di Oncologia Molecolare Milano Italy

Institute of Molecular Genetics of the Czech Academy of Sciences Prague Czech Republic

Pathology Department Complejo Hospitalario Universitario Insular Materno Infantil Las Palmas de Gran Canaria Spain

Universita' degli Studi di Milano Milano Italy

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Actionable cancer vulnerability due to translational arrest, p53 aggregation and ribosome biogenesis stress evoked by the disulfiram metabolite CuET