The SLX4 tumor suppressor is a scaffold that plays a pivotal role in several aspects of genome protection, including homologous recombination, interstrand DNA crosslink repair and the maintenance of common fragile sites and telomeres. Here, we unravel an unexpected direct interaction between SLX4 and the DNA helicase RTEL1, which, until now, were viewed as having independent and antagonistic functions. We identify cancer and Hoyeraal-Hreidarsson syndrome-associated mutations in SLX4 and RTEL1, respectively, that abolish SLX4-RTEL1 complex formation. We show that both proteins are recruited to nascent DNA, tightly co-localize with active RNA pol II, and that SLX4, in complex with RTEL1, promotes FANCD2/RNA pol II co-localization. Importantly, disrupting the SLX4-RTEL1 interaction leads to DNA replication defects in unstressed cells, which are rescued by inhibiting transcription. Our data demonstrate that SLX4 and RTEL1 interact to prevent replication-transcription conflicts and provide evidence that this is independent of the nuclease scaffold function of SLX4.

Centre de Recherche en Cancérologie de Marseille CRCM Inserm CNRS Aix Marseille Université Institut Paoli Calmettes Marseille France

CNRS UMR9019 Université Paris Saclay Equipe labellisée Ligue contre le Cancer Gustave Roussy Villejuif France

CNRS UMR9019 Université Paris Saclay Gustave Roussy Villejuif France

Department of Chemistry Faculty of Science Fukuoka University Fukuoka Japan

Inovarion Paris France

INSERM UMR 1163 Laboratory of Genome Dynamics in the Immune System Equipe Labellisée La Ligue contre le Cancer Paris Descartes Sorbonne Paris Cité University Imagine Institute Paris France

Institut Curie PSL Research University CNRS UMR3244 Paris France

Institute for Integrative Biology of the Cell CEA CNRS Univ Paris Sud Université Paris Saclay Gif sur Yvette cedex France

Institute of Molecular Genetics Prague Czech Republic

Sorbonne Université Muséum National d'Histoire Naturelle UMR CNRS 7590 IRD Institut de Minéralogie de Physique des Matériaux et de Cosmochimie IMPMC Paris France

Sorbonne Universités UPMC Univ Paris 06 CNRS UMR3244 Paris France

Erratum v

Nat Struct Mol Biol. 2020 Jun;27(6):604. doi: 10.1038/s41594-020-0447-z. PubMed

Erratum v

Nat Struct Mol Biol. 2020 Jun;27(6):603. doi: 10.1038/s41594-020-0448-y. PubMed

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Human senataxin is a bona fide R-loop resolving enzyme and transcription termination factor