biostudies-literatureSoler-Oliva MEMinisterio de Ciencia, Innovación y UniversidadesEuropean Research CouncilFundación de Investigación de la Universidad de SevillaUniversidad de SevillaAgencia Estatal de Investigacióne1012040https://www.ebi.ac.uk/biostudies/studies/S-EPMC12912698biostudies-literatureUnknown22(2)Transcription is essential for cellular function, but it can also lead to genetic instability, particularly through the formation of secondary structures such as R-loops, which consist of an RNA-DNA hybrid and a displaced DNA strand. Unscheduled R-loop accumulation is a major source of DNA damage and has been associated with several human diseases, including cancer. While multiple factors involved in RNA biogenesis, export, and chromatin remodeling play a role in preventing R-loop accumulation, the function of essential proteins in R-loop metabolism remains unexplored. Here, we performed a genetic screening in Saccharomyces cerevisiae using over 1200 temperature-sensitive mutants to identify novel proteins involved in the prevention of R-loop-associated genomic instability. Our results revPLoS geneticsAn essential gene screening identifies yeast Mot1 as a suppressor of R-loops and genome instability.PMC12912698PID2019-104270GB-I00/BMCVII PPIT-USPID2022-138251NB-I00ERC2014 AdG669898 TARLOOPFIUS22/0178PID2022-140466NB-I00Dominguez-Sierra RAAguilera ASoler-Oliva MEGaillard HfalseAn essential gene screening identifies yeast Mot1 as a suppressor of R-loops and genome instability.Transcription is essential for cellular function, but it can also lead to genetic instability, particularly through the formation of secondary structures such as R-loops, which consist of an RNA-DNA hybrid and a displaced DNA strand. Unscheduled R-loop accumulation is a major source of DNA damage and has been associated with several human diseases, including cancer. While multiple factors involved in RNA biogenesis, export, and chromatin remodeling play a role in preventing R-loop accumulation, the function of essential proteins in R-loop metabolism remains unexplored. Here, we performed a genetic screening in Saccharomyces cerevisiae using over 1200 temperature-sensitive mutants to identify novel proteins involved in the prevention of R-loop-associated genomic instability. Our results rev2026-01-01T00:00:00Z2026 Feb2026-07-16T00:37:11.409Z2026-07-09T10:26:11.115ZS-EPMC129126984166239710.1371/journal.pgen.1012040