In Vivo Occupancy of Mitochondrial Single-Stranded DNA Binding Protein Supports the Strand Displacement Mode of DNA Replication

Mitochondrial DNA (mtDNA) encodes for proteins required for oxidative phosphorylation, and mutations affecting the genome have been linked to a number of diseases as well as the natural ageing process in mammals. Human mtDNA is replicated by a molecular machinery that is distinct from the nuclear re...

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Autores principales: Miralles Fusté, Javier, Shi, Yonghong, Wanrooij, Sjoerd, Zhu, Xuefeng, Jemt, Elisabeth, Persson, Örjan, Sabouri, Nasim, Gustafsson, Claes M., Falkenberg, Maria
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4256270/
https://www.ncbi.nlm.nih.gov/pubmed/25474639
http://dx.doi.org/10.1371/journal.pgen.1004832
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author Miralles Fusté, Javier
Shi, Yonghong
Wanrooij, Sjoerd
Zhu, Xuefeng
Jemt, Elisabeth
Persson, Örjan
Sabouri, Nasim
Gustafsson, Claes M.
Falkenberg, Maria
author_facet Miralles Fusté, Javier
Shi, Yonghong
Wanrooij, Sjoerd
Zhu, Xuefeng
Jemt, Elisabeth
Persson, Örjan
Sabouri, Nasim
Gustafsson, Claes M.
Falkenberg, Maria
author_sort Miralles Fusté, Javier
collection PubMed
description Mitochondrial DNA (mtDNA) encodes for proteins required for oxidative phosphorylation, and mutations affecting the genome have been linked to a number of diseases as well as the natural ageing process in mammals. Human mtDNA is replicated by a molecular machinery that is distinct from the nuclear replisome, but there is still no consensus on the exact mode of mtDNA replication. We here demonstrate that the mitochondrial single-stranded DNA binding protein (mtSSB) directs origin specific initiation of mtDNA replication. MtSSB covers the parental heavy strand, which is displaced during mtDNA replication. MtSSB blocks primer synthesis on the displaced strand and restricts initiation of light-strand mtDNA synthesis to the specific origin of light-strand DNA synthesis (OriL). The in vivo occupancy profile of mtSSB displays a distinct pattern, with the highest levels of mtSSB close to the mitochondrial control region and with a gradual decline towards OriL. The pattern correlates with the replication products expected for the strand displacement mode of mtDNA synthesis, lending strong in vivo support for this debated model for mitochondrial DNA replication.
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spelling pubmed-42562702014-12-11 In Vivo Occupancy of Mitochondrial Single-Stranded DNA Binding Protein Supports the Strand Displacement Mode of DNA Replication Miralles Fusté, Javier Shi, Yonghong Wanrooij, Sjoerd Zhu, Xuefeng Jemt, Elisabeth Persson, Örjan Sabouri, Nasim Gustafsson, Claes M. Falkenberg, Maria PLoS Genet Research Article Mitochondrial DNA (mtDNA) encodes for proteins required for oxidative phosphorylation, and mutations affecting the genome have been linked to a number of diseases as well as the natural ageing process in mammals. Human mtDNA is replicated by a molecular machinery that is distinct from the nuclear replisome, but there is still no consensus on the exact mode of mtDNA replication. We here demonstrate that the mitochondrial single-stranded DNA binding protein (mtSSB) directs origin specific initiation of mtDNA replication. MtSSB covers the parental heavy strand, which is displaced during mtDNA replication. MtSSB blocks primer synthesis on the displaced strand and restricts initiation of light-strand mtDNA synthesis to the specific origin of light-strand DNA synthesis (OriL). The in vivo occupancy profile of mtSSB displays a distinct pattern, with the highest levels of mtSSB close to the mitochondrial control region and with a gradual decline towards OriL. The pattern correlates with the replication products expected for the strand displacement mode of mtDNA synthesis, lending strong in vivo support for this debated model for mitochondrial DNA replication. Public Library of Science 2014-12-04 /pmc/articles/PMC4256270/ /pubmed/25474639 http://dx.doi.org/10.1371/journal.pgen.1004832 Text en © 2014 Fusté 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
Miralles Fusté, Javier
Shi, Yonghong
Wanrooij, Sjoerd
Zhu, Xuefeng
Jemt, Elisabeth
Persson, Örjan
Sabouri, Nasim
Gustafsson, Claes M.
Falkenberg, Maria
In Vivo Occupancy of Mitochondrial Single-Stranded DNA Binding Protein Supports the Strand Displacement Mode of DNA Replication
title In Vivo Occupancy of Mitochondrial Single-Stranded DNA Binding Protein Supports the Strand Displacement Mode of DNA Replication
title_full In Vivo Occupancy of Mitochondrial Single-Stranded DNA Binding Protein Supports the Strand Displacement Mode of DNA Replication
title_fullStr In Vivo Occupancy of Mitochondrial Single-Stranded DNA Binding Protein Supports the Strand Displacement Mode of DNA Replication
title_full_unstemmed In Vivo Occupancy of Mitochondrial Single-Stranded DNA Binding Protein Supports the Strand Displacement Mode of DNA Replication
title_short In Vivo Occupancy of Mitochondrial Single-Stranded DNA Binding Protein Supports the Strand Displacement Mode of DNA Replication
title_sort in vivo occupancy of mitochondrial single-stranded dna binding protein supports the strand displacement mode of dna replication
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4256270/
https://www.ncbi.nlm.nih.gov/pubmed/25474639
http://dx.doi.org/10.1371/journal.pgen.1004832
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