title: The SET2-RPB1 interaction domain of human RECQ5 is important for transcription-associated genome stability.
abstract: The conserved RECQ5 DNA helicase is a tumor suppressor in mammalian cells. Defects in RECQ5 lead to the accumulation of spontaneous DNA double-stranded breaks (DSBs) during replication, despite the fact that these cells are proficient in DSB repair by homologous recombination (HR). The reason for this is unknown. Here, we demonstrate that these DSBs are linked to RNA polymerase II (RNAPII)-dependent transcription. In human RECQ5-depleted cells, active RNAPII accumulates on chromatin, and DNA breaks are associated with an RNAPII-dependent transcribed locus. Hence, transcription inhibition eliminates both active RNAPII and spontaneous DSB formation. In addition, the regulatory effect of RECQ5 on transcription and its interaction with RNAPII are enhanced in S-phase cells, supporting a role for RECQ5 in preventing transcription-associated DSBs during replication. Finally, we show that the SET2-RPB1 interaction (SRI) domain of human RECQ5 is important for suppressing spontaneous DSBs and the p53-dependent transcription stress response caused by the stalling of active RNAPII on DNA. Thus, our studies provide novel insights into a mechanism by which RECQ5 regulates the transcription machinery via its dynamic interaction with RNAPII, thereby preventing genome instability.
authors.affiliation: Department of Therapeutic Radiology, Yale University School of Medicine, 15 York St., New Haven, CT 06511, USA.
authors.forename: Min
authors.initials: M
authors.lastname: Li
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Xu
authors.forename: Yilun
authors.initials: Y
authors.lastname: Liu
mesh_terms: D002453:Cell Cycle; D002843:Chromatin; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D004261:DNA Replication; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D005434:Flow Cytometry; D015870:Gene Expression; D020868:Gene Silencing; D042822:Genomic Instability; D011495:Histone-Lysine N-Methyltransferase; D006801:Humans; D047468:Immunoprecipitation; D016133:Polymerase Chain Reaction; D054730:Protein Interaction Domains and Motifs; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering; D053484:RecQ Helicases / Q000737:chemistry* / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: D002843:Chromatin; C506732:RECQL5 protein, human; D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53; D011495:Histone-Lysine N-Methyltransferase; C521636:SETD2 protein, human; D012319:RNA Polymerase II; D012321:DNA-Directed RNA Polymerases; D053484:RecQ Helicases