Physical proximity of chromatin to nuclear pores prevents harmful R loop accumulation contributing to maintain genome stability

During transcription, the mRNA may hybridize with DNA, forming an R loop, which can be physiological or pathological, constituting in this case a source of genomic instability. To understand the mechanism by which eukaryotic cells prevent harmful R loops, we used human activation-induced cytidine deaminase (AID) to identify genes preventing R loops. A screening of 400 Saccharomyces cerevisiae selected strains deleted in nuclear genes revealed that cells lacking the Mlp1/2 nuclear basket proteins show AID-dependent genomic instability and replication defects that were suppressed by RNase H1 overexpression. Importantly, DNA–RNA hybrids accumulated at transcribed genes in mlp1/2 mutants, indicating that Mlp1/2 prevents R loops. Consistent with the Mlp1/2 role in gene gating to nuclear pores, artificial tethering to the nuclear periphery of a transcribed locus suppressed R loops in mlp1Δ cells. The same occurred in THO-deficient hpr1Δ cells. We conclude that proximity of transcribed chromatin to the nuclear pore helps restrain pathological R loops.Research was funded by the European Research Council (ERC2014 AdG669898 TARLOOP), the Spanish Ministry of Economy and Competitiveness (BFU2013-42918-P and BFU2016-75058-P), and the European Union (Fondo Europeo de Desarrollo Regional). F.G.-B. was the recipient of a predoctoral grant from the Spanish Ministry of Education, Culture and Sport (Formación de Profesorado Universitario).Peer Reviewe