Cargando…
Double Holliday Junctions are Intermediates of DNA Break Repair
Repair of DNA double-strand-breaks (DSBs) by homologous recombination is crucial for cell proliferation and tumor suppression. However, despite its importance, the molecular intermediates of mitotic DSB-repair remain undefined. The double Holliday Junction (dHJ), presupposed to be the central interm...
Autores principales: | , , , , |
---|---|
Formato: | Texto |
Lenguaje: | English |
Publicado: |
2010
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2851831/ https://www.ncbi.nlm.nih.gov/pubmed/20348905 http://dx.doi.org/10.1038/nature08868 |
_version_ | 1782179905075675136 |
---|---|
author | Bzymek, Malgorzata Thayer, Nathaniel H. Oh, Steve D. Kleckner, Nancy Hunter, Neil |
author_facet | Bzymek, Malgorzata Thayer, Nathaniel H. Oh, Steve D. Kleckner, Nancy Hunter, Neil |
author_sort | Bzymek, Malgorzata |
collection | PubMed |
description | Repair of DNA double-strand-breaks (DSBs) by homologous recombination is crucial for cell proliferation and tumor suppression. However, despite its importance, the molecular intermediates of mitotic DSB-repair remain undefined. The double Holliday Junction (dHJ), presupposed to be the central intermediate for more than 25 years1, has only been identified during meiotic recombination2. Moreover, evidence has accumulated for alternative, dHJ-independent mechanisms3–6, raising the possibility that dHJs are not formed during DSB-repair in mitotically cycling cells. Here we identify intermediates of DSB-repair using a budding yeast assay system designed to mimic physiological DSB repair. This system utilizes diploid cells and provides the possibility for allelic recombination either between sister-chromatids or between homologs, as well as direct comparison with meiotic recombination at the same locus. In mitotically cycling cells, we detect inter-homolog Joint Molecule (JM) intermediates whose size and strand-composition are identical to the canonical dHJ structures observed in meiosis2. However, in contrast to meiosis, JMs between sister chromatids form in preference to those between homologs. Moreover, JMs appear to represent a minor pathway of DSB repair in mitotic cells, being detected at ~10-fold lower levels (per DSB) than during meiotic recombination. Thus, although dHJs are identified as intermediates of DSB-promoted recombination in both mitotic and meiotic cells, their formation is distinctly regulated according to the specific dictates of the two cellular programs. |
format | Text |
id | pubmed-2851831 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
record_format | MEDLINE/PubMed |
spelling | pubmed-28518312010-10-08 Double Holliday Junctions are Intermediates of DNA Break Repair Bzymek, Malgorzata Thayer, Nathaniel H. Oh, Steve D. Kleckner, Nancy Hunter, Neil Nature Article Repair of DNA double-strand-breaks (DSBs) by homologous recombination is crucial for cell proliferation and tumor suppression. However, despite its importance, the molecular intermediates of mitotic DSB-repair remain undefined. The double Holliday Junction (dHJ), presupposed to be the central intermediate for more than 25 years1, has only been identified during meiotic recombination2. Moreover, evidence has accumulated for alternative, dHJ-independent mechanisms3–6, raising the possibility that dHJs are not formed during DSB-repair in mitotically cycling cells. Here we identify intermediates of DSB-repair using a budding yeast assay system designed to mimic physiological DSB repair. This system utilizes diploid cells and provides the possibility for allelic recombination either between sister-chromatids or between homologs, as well as direct comparison with meiotic recombination at the same locus. In mitotically cycling cells, we detect inter-homolog Joint Molecule (JM) intermediates whose size and strand-composition are identical to the canonical dHJ structures observed in meiosis2. However, in contrast to meiosis, JMs between sister chromatids form in preference to those between homologs. Moreover, JMs appear to represent a minor pathway of DSB repair in mitotic cells, being detected at ~10-fold lower levels (per DSB) than during meiotic recombination. Thus, although dHJs are identified as intermediates of DSB-promoted recombination in both mitotic and meiotic cells, their formation is distinctly regulated according to the specific dictates of the two cellular programs. 2010-03-28 2010-04-08 /pmc/articles/PMC2851831/ /pubmed/20348905 http://dx.doi.org/10.1038/nature08868 Text en Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms |
spellingShingle | Article Bzymek, Malgorzata Thayer, Nathaniel H. Oh, Steve D. Kleckner, Nancy Hunter, Neil Double Holliday Junctions are Intermediates of DNA Break Repair |
title | Double Holliday Junctions are Intermediates of DNA Break Repair |
title_full | Double Holliday Junctions are Intermediates of DNA Break Repair |
title_fullStr | Double Holliday Junctions are Intermediates of DNA Break Repair |
title_full_unstemmed | Double Holliday Junctions are Intermediates of DNA Break Repair |
title_short | Double Holliday Junctions are Intermediates of DNA Break Repair |
title_sort | double holliday junctions are intermediates of dna break repair |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2851831/ https://www.ncbi.nlm.nih.gov/pubmed/20348905 http://dx.doi.org/10.1038/nature08868 |
work_keys_str_mv | AT bzymekmalgorzata doublehollidayjunctionsareintermediatesofdnabreakrepair AT thayernathanielh doublehollidayjunctionsareintermediatesofdnabreakrepair AT ohsteved doublehollidayjunctionsareintermediatesofdnabreakrepair AT klecknernancy doublehollidayjunctionsareintermediatesofdnabreakrepair AT hunterneil doublehollidayjunctionsareintermediatesofdnabreakrepair |