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Rtt105 regulates RPA function by configurationally stapling the flexible domains

Replication Protein A (RPA) is a heterotrimeric complex that binds to single-stranded DNA (ssDNA) and recruits over three dozen RPA-interacting proteins to coordinate multiple aspects of DNA metabolism including DNA replication, repair, and recombination. Rtt105 is a molecular chaperone that regulat...

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Autores principales: Kuppa, Sahiti, Deveryshetty, Jaigeeth, Chadda, Rahul, Mattice, Jenna R., Pokhrel, Nilisha, Kaushik, Vikas, Patterson, Angela, Dhingra, Nalini, Pangeni, Sushil, Sadauskas, Marisa K., Shiekh, Sajad, Balci, Hamza, Ha, Taekjip, Zhao, Xiaolan, Bothner, Brian, Antony, Edwin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9440123/
https://www.ncbi.nlm.nih.gov/pubmed/36056028
http://dx.doi.org/10.1038/s41467-022-32860-6
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author Kuppa, Sahiti
Deveryshetty, Jaigeeth
Chadda, Rahul
Mattice, Jenna R.
Pokhrel, Nilisha
Kaushik, Vikas
Patterson, Angela
Dhingra, Nalini
Pangeni, Sushil
Sadauskas, Marisa K.
Shiekh, Sajad
Balci, Hamza
Ha, Taekjip
Zhao, Xiaolan
Bothner, Brian
Antony, Edwin
author_facet Kuppa, Sahiti
Deveryshetty, Jaigeeth
Chadda, Rahul
Mattice, Jenna R.
Pokhrel, Nilisha
Kaushik, Vikas
Patterson, Angela
Dhingra, Nalini
Pangeni, Sushil
Sadauskas, Marisa K.
Shiekh, Sajad
Balci, Hamza
Ha, Taekjip
Zhao, Xiaolan
Bothner, Brian
Antony, Edwin
author_sort Kuppa, Sahiti
collection PubMed
description Replication Protein A (RPA) is a heterotrimeric complex that binds to single-stranded DNA (ssDNA) and recruits over three dozen RPA-interacting proteins to coordinate multiple aspects of DNA metabolism including DNA replication, repair, and recombination. Rtt105 is a molecular chaperone that regulates nuclear localization of RPA. Here, we show that Rtt105 binds to multiple DNA binding and protein-interaction domains of RPA and configurationally staples the complex. In the absence of ssDNA, Rtt105 inhibits RPA binding to Rad52, thus preventing spurious binding to RPA-interacting proteins. When ssDNA is available, Rtt105 promotes formation of high-density RPA nucleoprotein filaments and dissociates during this process. Free Rtt105 further stabilizes the RPA-ssDNA filaments by inhibiting the facilitated exchange activity of RPA. Collectively, our data suggest that Rtt105 sequesters free RPA in the nucleus to prevent untimely binding to RPA-interacting proteins, while stabilizing RPA-ssDNA filaments at DNA lesion sites.
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spelling pubmed-94401232022-09-04 Rtt105 regulates RPA function by configurationally stapling the flexible domains Kuppa, Sahiti Deveryshetty, Jaigeeth Chadda, Rahul Mattice, Jenna R. Pokhrel, Nilisha Kaushik, Vikas Patterson, Angela Dhingra, Nalini Pangeni, Sushil Sadauskas, Marisa K. Shiekh, Sajad Balci, Hamza Ha, Taekjip Zhao, Xiaolan Bothner, Brian Antony, Edwin Nat Commun Article Replication Protein A (RPA) is a heterotrimeric complex that binds to single-stranded DNA (ssDNA) and recruits over three dozen RPA-interacting proteins to coordinate multiple aspects of DNA metabolism including DNA replication, repair, and recombination. Rtt105 is a molecular chaperone that regulates nuclear localization of RPA. Here, we show that Rtt105 binds to multiple DNA binding and protein-interaction domains of RPA and configurationally staples the complex. In the absence of ssDNA, Rtt105 inhibits RPA binding to Rad52, thus preventing spurious binding to RPA-interacting proteins. When ssDNA is available, Rtt105 promotes formation of high-density RPA nucleoprotein filaments and dissociates during this process. Free Rtt105 further stabilizes the RPA-ssDNA filaments by inhibiting the facilitated exchange activity of RPA. Collectively, our data suggest that Rtt105 sequesters free RPA in the nucleus to prevent untimely binding to RPA-interacting proteins, while stabilizing RPA-ssDNA filaments at DNA lesion sites. Nature Publishing Group UK 2022-09-02 /pmc/articles/PMC9440123/ /pubmed/36056028 http://dx.doi.org/10.1038/s41467-022-32860-6 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Kuppa, Sahiti
Deveryshetty, Jaigeeth
Chadda, Rahul
Mattice, Jenna R.
Pokhrel, Nilisha
Kaushik, Vikas
Patterson, Angela
Dhingra, Nalini
Pangeni, Sushil
Sadauskas, Marisa K.
Shiekh, Sajad
Balci, Hamza
Ha, Taekjip
Zhao, Xiaolan
Bothner, Brian
Antony, Edwin
Rtt105 regulates RPA function by configurationally stapling the flexible domains
title Rtt105 regulates RPA function by configurationally stapling the flexible domains
title_full Rtt105 regulates RPA function by configurationally stapling the flexible domains
title_fullStr Rtt105 regulates RPA function by configurationally stapling the flexible domains
title_full_unstemmed Rtt105 regulates RPA function by configurationally stapling the flexible domains
title_short Rtt105 regulates RPA function by configurationally stapling the flexible domains
title_sort rtt105 regulates rpa function by configurationally stapling the flexible domains
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9440123/
https://www.ncbi.nlm.nih.gov/pubmed/36056028
http://dx.doi.org/10.1038/s41467-022-32860-6
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