Yeast Assay Highlights the Intrinsic Genomic Instability of Human PML Intron 6 over Intron 3 and the Role of Replication Fork Proteins

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 mech...

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Autores principales: Chanet, Roland, Kienda, Guy, Heneman-Masurel, Amélie, Vernis, Laurence, Cassinat, Bruno, Guardiola, Philippe, Fenaux, Pierre, Chomienne, Christine, Huang, Meng-Er
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4460018/
https://www.ncbi.nlm.nih.gov/pubmed/26053431
http://dx.doi.org/10.1371/journal.pone.0129222
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author Chanet, Roland
Kienda, Guy
Heneman-Masurel, Amélie
Vernis, Laurence
Cassinat, Bruno
Guardiola, Philippe
Fenaux, Pierre
Chomienne, Christine
Huang, Meng-Er
author_facet Chanet, Roland
Kienda, Guy
Heneman-Masurel, Amélie
Vernis, Laurence
Cassinat, Bruno
Guardiola, Philippe
Fenaux, Pierre
Chomienne, Christine
Huang, Meng-Er
author_sort Chanet, Roland
collection PubMed
description 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.
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spelling pubmed-44600182015-06-16 Yeast Assay Highlights the Intrinsic Genomic Instability of Human PML Intron 6 over Intron 3 and the Role of Replication Fork Proteins Chanet, Roland Kienda, Guy Heneman-Masurel, Amélie Vernis, Laurence Cassinat, Bruno Guardiola, Philippe Fenaux, Pierre Chomienne, Christine Huang, Meng-Er PLoS One Research Article 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. Public Library of Science 2015-06-08 /pmc/articles/PMC4460018/ /pubmed/26053431 http://dx.doi.org/10.1371/journal.pone.0129222 Text en © 2015 Chanet et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Chanet, Roland
Kienda, Guy
Heneman-Masurel, Amélie
Vernis, Laurence
Cassinat, Bruno
Guardiola, Philippe
Fenaux, Pierre
Chomienne, Christine
Huang, Meng-Er
Yeast Assay Highlights the Intrinsic Genomic Instability of Human PML Intron 6 over Intron 3 and the Role of Replication Fork Proteins
title Yeast Assay Highlights the Intrinsic Genomic Instability of Human PML Intron 6 over Intron 3 and the Role of Replication Fork Proteins
title_full Yeast Assay Highlights the Intrinsic Genomic Instability of Human PML Intron 6 over Intron 3 and the Role of Replication Fork Proteins
title_fullStr Yeast Assay Highlights the Intrinsic Genomic Instability of Human PML Intron 6 over Intron 3 and the Role of Replication Fork Proteins
title_full_unstemmed Yeast Assay Highlights the Intrinsic Genomic Instability of Human PML Intron 6 over Intron 3 and the Role of Replication Fork Proteins
title_short Yeast Assay Highlights the Intrinsic Genomic Instability of Human PML Intron 6 over Intron 3 and the Role of Replication Fork Proteins
title_sort yeast assay highlights the intrinsic genomic instability of human pml intron 6 over intron 3 and the role of replication fork proteins
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4460018/
https://www.ncbi.nlm.nih.gov/pubmed/26053431
http://dx.doi.org/10.1371/journal.pone.0129222
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