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Generation of double Holliday junction DNAs and their dissolution/resolution within a chromatin context
Four-way DNA intermediates, also known as Holliday junctions (HJs), are formed during homologous recombination and DNA repair, and their resolution is necessary for proper chromosome segregation. To facilitate the biochemical analysis of HJ processing, we developed a method involving DNAzyme self-cl...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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National Academy of Sciences
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9170140/ https://www.ncbi.nlm.nih.gov/pubmed/35452329 http://dx.doi.org/10.1073/pnas.2123420119 |
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author | Ho, Han N. West, Stephen C. |
author_facet | Ho, Han N. West, Stephen C. |
author_sort | Ho, Han N. |
collection | PubMed |
description | Four-way DNA intermediates, also known as Holliday junctions (HJs), are formed during homologous recombination and DNA repair, and their resolution is necessary for proper chromosome segregation. To facilitate the biochemical analysis of HJ processing, we developed a method involving DNAzyme self-cleavage to generate 1.8-kb DNA molecules containing either single (sHJ) or double Holliday junctions (dHJs). We show that dHJ DNAs (referred to as HoJo DNAs) are dissolved by the human BLM–TopIIIα–RMI1–RMI2 complex to form two noncrossover products. However, structure-selective endonucleases (human GEN1 and SMX complex) resolve DNA containing single or double HJs to yield a mixture of crossover and noncrossover products. Finally, we demonstrate that chromatin inhibits the resolution of the double HJ by GEN or SMX while allowing BTRR-mediated dissolution. |
format | Online Article Text |
id | pubmed-9170140 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | National Academy of Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-91701402022-06-07 Generation of double Holliday junction DNAs and their dissolution/resolution within a chromatin context Ho, Han N. West, Stephen C. Proc Natl Acad Sci U S A Biological Sciences Four-way DNA intermediates, also known as Holliday junctions (HJs), are formed during homologous recombination and DNA repair, and their resolution is necessary for proper chromosome segregation. To facilitate the biochemical analysis of HJ processing, we developed a method involving DNAzyme self-cleavage to generate 1.8-kb DNA molecules containing either single (sHJ) or double Holliday junctions (dHJs). We show that dHJ DNAs (referred to as HoJo DNAs) are dissolved by the human BLM–TopIIIα–RMI1–RMI2 complex to form two noncrossover products. However, structure-selective endonucleases (human GEN1 and SMX complex) resolve DNA containing single or double HJs to yield a mixture of crossover and noncrossover products. Finally, we demonstrate that chromatin inhibits the resolution of the double HJ by GEN or SMX while allowing BTRR-mediated dissolution. National Academy of Sciences 2022-04-22 2022-05-03 /pmc/articles/PMC9170140/ /pubmed/35452329 http://dx.doi.org/10.1073/pnas.2123420119 Text en Copyright © 2022 the Author(s). Published by PNAS https://creativecommons.org/licenses/by/4.0/This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Biological Sciences Ho, Han N. West, Stephen C. Generation of double Holliday junction DNAs and their dissolution/resolution within a chromatin context |
title | Generation of double Holliday junction DNAs and their dissolution/resolution within a chromatin context |
title_full | Generation of double Holliday junction DNAs and their dissolution/resolution within a chromatin context |
title_fullStr | Generation of double Holliday junction DNAs and their dissolution/resolution within a chromatin context |
title_full_unstemmed | Generation of double Holliday junction DNAs and their dissolution/resolution within a chromatin context |
title_short | Generation of double Holliday junction DNAs and their dissolution/resolution within a chromatin context |
title_sort | generation of double holliday junction dnas and their dissolution/resolution within a chromatin context |
topic | Biological Sciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9170140/ https://www.ncbi.nlm.nih.gov/pubmed/35452329 http://dx.doi.org/10.1073/pnas.2123420119 |
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