title: Yeast Assay Highlights the Intrinsic Genomic Instability of Human PML Intron 6 over Intron 3 and the Role of Replication Fork Proteins.
abstract: Human acute promyelocytic leukemia (APL) is characterized by a specific balanced translocation t(15;17)(q22;q21) involving the PML and RARA genes. In both de novo and therapy-related APL, the most frequent PML breakpoints are located within intron 6, and less frequently in intron 3; the precise mechanisms by which these breakpoints arise and preferentially in PML intron 6 remain unsolved. To investigate the intrinsic properties of the PML intron sequences in vivo, we designed Saccharomyces cerevisiae strains containing human PML intron 6 or intron 3 sequences inserted in yeast chromosome V and measured gross chromosomal rearrangements (GCR). This approach provided evidence that intron 6 had a superior instability over intron 3 due to an intrinsic property of the sequence and identified the 3' end of intron 6 as the most susceptible to break. Using yeast strains invalidated for genes that control DNA replication, we show that this differential instability depended at least upon Rrm3, a DNA helicase, and Mrc1, the human claspin homolog. GCR induction by hydrogen peroxide, a general genotoxic agent, was also dependent on genetic context. We conclude that: 1) this yeast system provides an alternative approach to study in detail the properties of human sequences in a genetically controlled situation and 2) the different susceptibility to produce DNA breaks in intron 6 versus intron 3 of the human PML gene is likely due to an intrinsic property of the sequence and is under replication fork genetic control.
authors.affiliation: UMR3348 "Genotoxic Stress and Cancer", Centre National de la Recherche Scientifique, Orsay, France; Institut Curie, Centre de Recherche, Orsay, France.
authors.forename: Roland
authors.initials: R
authors.lastname: Chanet
authors.affiliation: UMR3348 "Genotoxic Stress and Cancer", Centre National de la Recherche Scientifique, Orsay, France; Institut Curie, Centre de Recherche, Orsay, France.
authors.forename: Guy
authors.initials: G
authors.lastname: Kienda
authors.affiliation: UMR3348 "Genotoxic Stress and Cancer", Centre National de la Recherche Scientifique, Orsay, France; Institut Curie, Centre de Recherche, Orsay, France.
authors.forename: Amélie
authors.initials: A
authors.lastname: Heneman-Masurel
authors.affiliation: UMR3348 "Genotoxic Stress and Cancer", Centre National de la Recherche Scientifique, Orsay, France; Institut Curie, Centre de Recherche, Orsay, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Vernis
authors.affiliation: AP-HP, Hôpital Saint-Louis, Paris, France; Inserm UMRS-1131, Hôpital Saint-Louis, Université Paris Diderot, Paris, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Cassinat
authors.affiliation: Plateforme SNP, Transcriptome & Epigénomique, Centre Hospitalier Universitaire, Angers, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Guardiola
authors.affiliation: AP-HP, Hôpital Saint-Louis, Paris, France; Inserm UMRS-1131, Hôpital Saint-Louis, Université Paris Diderot, Paris, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Fenaux
authors.affiliation: AP-HP, Hôpital Saint-Louis, Paris, France; Inserm UMRS-1131, Hôpital Saint-Louis, Université Paris Diderot, Paris, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Chomienne
authors.affiliation: UMR3348 "Genotoxic Stress and Cancer", Centre National de la Recherche Scientifique, Orsay, France; Institut Curie, Centre de Recherche, Orsay, France.
authors.forename: Meng-Er
authors.initials: ME
authors.lastname: Huang
mesh_terms: D056905:Chromosome Breakpoints; D002874:Chromosome Mapping; D053960:DNA Breaks / Q000187:drug effects; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000378:metabolism*; D023061:Gene Order; D056426:Genetic Loci; D042822:Genomic Instability; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007438:Introns; D015473:Leukemia, Promyelocytic, Acute / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics*; D000071156:Promyelocytic Leukemia Protein; D018168:Receptors, Retinoic Acid / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics*; D014178:Translocation, Genetic; D025521:Tumor Suppressor Proteins / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D018168:Receptors, Retinoic Acid; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C070576:PML protein, human; D006861:Hydrogen Peroxide