title: BRCA2 prevents R-loop accumulation and associates with TREX-2 mRNA export factor PCID2.
abstract: Genome instability is central to ageing, cancer and other diseases. It is not only proteins involved in DNA replication or the DNA damage response (DDR) that are important for maintaining genome integrity: from yeast to higher eukaryotes, mutations in genes involved in pre-mRNA splicing and in the biogenesis and export of messenger ribonucleoprotein (mRNP) also induce DNA damage and genome instability. This instability is frequently mediated by R-loops formed by DNA-RNA hybrids and a displaced single-stranded DNA. Here we show that the human TREX-2 complex, which is involved in mRNP biogenesis and export, prevents genome instability as determined by the accumulation of γ-H2AX (Ser-139 phosphorylated histone H2AX) and 53BP1 foci and single-cell electrophoresis in cells depleted of the TREX-2 subunits PCID2, GANP and DSS1. We show that the BRCA2 repair factor, which binds to DSS1, also associates with PCID2 in the cell. The use of an enhanced green fluorescent protein-tagged hybrid-binding domain of RNase H1 and the S9.6 antibody did not detect R-loops in TREX-2-depleted cells, but did detect the accumulation of R-loops in BRCA2-depleted cells. The results indicate that R-loops are frequently formed in cells and that BRCA2 is required for their processing. This link between BRCA2 and RNA-mediated genome instability indicates that R-loops may be a chief source of replication stress and cancer-associated instability.
authors.affiliation: Centro Andaluz de Biología Molecular y Medicina Regenerativa CABIMER, Universidad de Sevilla, Avenida Américo Vespucio s/n, 41092 Seville, Spain.
authors.forename: Vaibhav
authors.initials: V
authors.lastname: Bhatia
authors.affiliation: Centro Andaluz de Biología Molecular y Medicina Regenerativa CABIMER, Universidad de Sevilla, Avenida Américo Vespucio s/n, 41092 Seville, Spain.
authors.forename: Sonia I
authors.initials: SI
authors.lastname: Barroso
authors.affiliation: Centro Andaluz de Biología Molecular y Medicina Regenerativa CABIMER, Universidad de Sevilla, Avenida Américo Vespucio s/n, 41092 Seville, Spain.
authors.forename: María L
authors.initials: ML
authors.lastname: García-Rubio
authors.affiliation: Centro Andaluz de Biología Molecular y Medicina Regenerativa CABIMER, Universidad de Sevilla, Avenida Américo Vespucio s/n, 41092 Seville, Spain.
authors.forename: Emanuela
authors.initials: E
authors.lastname: Tumini
authors.affiliation: Centro Andaluz de Biología Molecular y Medicina Regenerativa CABIMER, Universidad de Sevilla, Avenida Américo Vespucio s/n, 41092 Seville, Spain.
authors.forename: Emilia
authors.initials: E
authors.lastname: Herrera-Moyano
authors.affiliation: Centro Andaluz de Biología Molecular y Medicina Regenerativa CABIMER, Universidad de Sevilla, Avenida Américo Vespucio s/n, 41092 Seville, Spain.
authors.forename: Andrés
authors.initials: A
authors.lastname: Aguilera
mesh_terms: D000123:Acetyltransferases / Q000378:metabolism; D024682:BRCA2 Protein / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D004249:DNA Damage; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism*; D005090:Exodeoxyribonucleases / Q000737:chemistry / Q000172:deficiency / Q000378:metabolism*; D042822:Genomic Instability; D006657:Histones / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D010750:Phosphoproteins / Q000737:chemistry / Q000172:deficiency / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D011485:Protein Binding; D012313:RNA / Q000737:chemistry / Q000378:metabolism*; D034443:RNA Transport; D016914:Ribonuclease H / Q000737:chemistry; D012261:Ribonucleoproteins / Q000096:biosynthesis / Q000378:metabolism
chemical_list: D024682:BRCA2 Protein; D004277:DNA, Single-Stranded; C468783:H2AX protein, human; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; C586854:PCID2 protein, human; D010750:Phosphoproteins; D012261:Ribonucleoproteins; C484606:SEM1 protein, human; C026452:messenger ribonucleoprotein; D012313:RNA; D000123:Acetyltransferases; C406825:MCM3AP protein, human; D005090:Exodeoxyribonucleases; C486104:TREX2 protein, human; D016914:Ribonuclease H; C078856:ribonuclease HI; D046988:Proteasome Endopeptidase Complex