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Recycling of single-stranded DNA-binding protein by the bacterial replisome

Single-stranded DNA-binding proteins (SSBs) support DNA replication by protecting single-stranded DNA from nucleolytic attack, preventing intra-strand pairing events and playing many other regulatory roles within the replisome. Recent developments in single-molecule approaches have led to a revised...

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Autores principales: Spenkelink, Lisanne M, Lewis, Jacob S, Jergic, Slobodan, Xu, Zhi-Qiang, Robinson, Andrew, Dixon, Nicholas E, van Oijen, Antoine M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6486552/
https://www.ncbi.nlm.nih.gov/pubmed/30767010
http://dx.doi.org/10.1093/nar/gkz090
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author Spenkelink, Lisanne M
Lewis, Jacob S
Jergic, Slobodan
Xu, Zhi-Qiang
Robinson, Andrew
Dixon, Nicholas E
van Oijen, Antoine M
author_facet Spenkelink, Lisanne M
Lewis, Jacob S
Jergic, Slobodan
Xu, Zhi-Qiang
Robinson, Andrew
Dixon, Nicholas E
van Oijen, Antoine M
author_sort Spenkelink, Lisanne M
collection PubMed
description Single-stranded DNA-binding proteins (SSBs) support DNA replication by protecting single-stranded DNA from nucleolytic attack, preventing intra-strand pairing events and playing many other regulatory roles within the replisome. Recent developments in single-molecule approaches have led to a revised picture of the replisome that is much more complex in how it retains or recycles protein components. Here, we visualize how an in vitro reconstituted Escherichia coli replisome recruits SSB by relying on two different molecular mechanisms. Not only does it recruit new SSB molecules from solution to coat newly formed single-stranded DNA on the lagging strand, but it also internally recycles SSB from one Okazaki fragment to the next. We show that this internal transfer mechanism is balanced against recruitment from solution in a manner that is concentration dependent. By visualizing SSB dynamics in live cells, we show that both internal transfer and external exchange mechanisms are physiologically relevant.
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spelling pubmed-64865522019-05-01 Recycling of single-stranded DNA-binding protein by the bacterial replisome Spenkelink, Lisanne M Lewis, Jacob S Jergic, Slobodan Xu, Zhi-Qiang Robinson, Andrew Dixon, Nicholas E van Oijen, Antoine M Nucleic Acids Res Genome Integrity, Repair and Replication Single-stranded DNA-binding proteins (SSBs) support DNA replication by protecting single-stranded DNA from nucleolytic attack, preventing intra-strand pairing events and playing many other regulatory roles within the replisome. Recent developments in single-molecule approaches have led to a revised picture of the replisome that is much more complex in how it retains or recycles protein components. Here, we visualize how an in vitro reconstituted Escherichia coli replisome recruits SSB by relying on two different molecular mechanisms. Not only does it recruit new SSB molecules from solution to coat newly formed single-stranded DNA on the lagging strand, but it also internally recycles SSB from one Okazaki fragment to the next. We show that this internal transfer mechanism is balanced against recruitment from solution in a manner that is concentration dependent. By visualizing SSB dynamics in live cells, we show that both internal transfer and external exchange mechanisms are physiologically relevant. Oxford University Press 2019-05-07 2019-02-15 /pmc/articles/PMC6486552/ /pubmed/30767010 http://dx.doi.org/10.1093/nar/gkz090 Text en © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Genome Integrity, Repair and Replication
Spenkelink, Lisanne M
Lewis, Jacob S
Jergic, Slobodan
Xu, Zhi-Qiang
Robinson, Andrew
Dixon, Nicholas E
van Oijen, Antoine M
Recycling of single-stranded DNA-binding protein by the bacterial replisome
title Recycling of single-stranded DNA-binding protein by the bacterial replisome
title_full Recycling of single-stranded DNA-binding protein by the bacterial replisome
title_fullStr Recycling of single-stranded DNA-binding protein by the bacterial replisome
title_full_unstemmed Recycling of single-stranded DNA-binding protein by the bacterial replisome
title_short Recycling of single-stranded DNA-binding protein by the bacterial replisome
title_sort recycling of single-stranded dna-binding protein by the bacterial replisome
topic Genome Integrity, Repair and Replication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6486552/
https://www.ncbi.nlm.nih.gov/pubmed/30767010
http://dx.doi.org/10.1093/nar/gkz090
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