Phosphorylation-dependent WRN-RPA interaction promotes recovery of stalled forks at secondary DNA structure
Abstract The WRN protein is vital for managing perturbed replication forks. Replication Protein A strongly enhances WRN helicase activity in specific in vitro assays. However, the in vivo significance of RPA binding to WRN has largely remained unexplored. We identify several conserved phosphorylatio...
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-55958-z |
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| author | Alessandro Noto Pasquale Valenzisi Flavia Di Feo Federica Fratini Tomasz Kulikowicz Joshua A. Sommers Benedetta Perdichizzi Maurizio Semproni Valentina Palermo Marco Crescenzi Robert M. Brosh Jr Annapaola Franchitto Pietro Pichierri |
| author_facet | Alessandro Noto Pasquale Valenzisi Flavia Di Feo Federica Fratini Tomasz Kulikowicz Joshua A. Sommers Benedetta Perdichizzi Maurizio Semproni Valentina Palermo Marco Crescenzi Robert M. Brosh Jr Annapaola Franchitto Pietro Pichierri |
| author_sort | Alessandro Noto |
| collection | DOAJ |
| description | Abstract The WRN protein is vital for managing perturbed replication forks. Replication Protein A strongly enhances WRN helicase activity in specific in vitro assays. However, the in vivo significance of RPA binding to WRN has largely remained unexplored. We identify several conserved phosphorylation sites in the acidic domain of WRN targeted by Casein Kinase 2. These phosphorylation sites are crucial for WRN-RPA interaction. Using an unphosphorylable WRN mutant, which lacks the ability to bind RPA, we determine that the WRN-RPA complex plays a critical role in fork recovery after replication stress countering the persistence of G4 structures after fork stalling. However, the interaction between WRN and RPA is not necessary for the processing of replication forks when they collapse. The absence of WRN-RPA binding hampers fork recovery, causing single-strand DNA gaps, enlarged by MRE11, and triggering MUS81-dependent double-strand breaks, which require repair by RAD51 to prevent excessive DNA damage. |
| format | Article |
| id | doaj-art-8af04971292747afb86a92dbe2841f9f |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-8af04971292747afb86a92dbe2841f9f2025-02-02T12:32:05ZengNature PortfolioNature Communications2041-17232025-01-0116112110.1038/s41467-025-55958-zPhosphorylation-dependent WRN-RPA interaction promotes recovery of stalled forks at secondary DNA structureAlessandro Noto0Pasquale Valenzisi1Flavia Di Feo2Federica Fratini3Tomasz Kulikowicz4Joshua A. Sommers5Benedetta Perdichizzi6Maurizio Semproni7Valentina Palermo8Marco Crescenzi9Robert M. Brosh Jr10Annapaola Franchitto11Pietro Pichierri12Mechanisms, Biomarkers and Models Section – Genome Stability Group, Department of Environment and Health, Istituto Superiore di SanitàMechanisms, Biomarkers and Models Section – Genome Stability Group, Department of Environment and Health, Istituto Superiore di SanitàMechanisms, Biomarkers and Models Section – Genome Stability Group, Department of Environment and Health, Istituto Superiore di SanitàCore Facilities Technical-Scientific Service – Mass Spectrometry Unit, Istituto Superiore di SanitàHelicases and Genomic Integrity Section, Translational Gerontology Branch, National Institute on Aging, NIHHelicases and Genomic Integrity Section, Translational Gerontology Branch, National Institute on Aging, NIHMechanisms, Biomarkers and Models Section – Genome Stability Group, Department of Environment and Health, Istituto Superiore di SanitàMechanisms, Biomarkers and Models Section – Genome Stability Group, Department of Environment and Health, Istituto Superiore di SanitàMechanisms, Biomarkers and Models Section – Genome Stability Group, Department of Environment and Health, Istituto Superiore di SanitàCore Facilities Technical-Scientific Service – Mass Spectrometry Unit, Istituto Superiore di SanitàHelicases and Genomic Integrity Section, Translational Gerontology Branch, National Institute on Aging, NIHMechanisms, Biomarkers and Models Section – Genome Stability Group, Department of Environment and Health, Istituto Superiore di SanitàMechanisms, Biomarkers and Models Section – Genome Stability Group, Department of Environment and Health, Istituto Superiore di SanitàAbstract The WRN protein is vital for managing perturbed replication forks. Replication Protein A strongly enhances WRN helicase activity in specific in vitro assays. However, the in vivo significance of RPA binding to WRN has largely remained unexplored. We identify several conserved phosphorylation sites in the acidic domain of WRN targeted by Casein Kinase 2. These phosphorylation sites are crucial for WRN-RPA interaction. Using an unphosphorylable WRN mutant, which lacks the ability to bind RPA, we determine that the WRN-RPA complex plays a critical role in fork recovery after replication stress countering the persistence of G4 structures after fork stalling. However, the interaction between WRN and RPA is not necessary for the processing of replication forks when they collapse. The absence of WRN-RPA binding hampers fork recovery, causing single-strand DNA gaps, enlarged by MRE11, and triggering MUS81-dependent double-strand breaks, which require repair by RAD51 to prevent excessive DNA damage.https://doi.org/10.1038/s41467-025-55958-z |
| spellingShingle | Alessandro Noto Pasquale Valenzisi Flavia Di Feo Federica Fratini Tomasz Kulikowicz Joshua A. Sommers Benedetta Perdichizzi Maurizio Semproni Valentina Palermo Marco Crescenzi Robert M. Brosh Jr Annapaola Franchitto Pietro Pichierri Phosphorylation-dependent WRN-RPA interaction promotes recovery of stalled forks at secondary DNA structure Nature Communications |
| title | Phosphorylation-dependent WRN-RPA interaction promotes recovery of stalled forks at secondary DNA structure |
| title_full | Phosphorylation-dependent WRN-RPA interaction promotes recovery of stalled forks at secondary DNA structure |
| title_fullStr | Phosphorylation-dependent WRN-RPA interaction promotes recovery of stalled forks at secondary DNA structure |
| title_full_unstemmed | Phosphorylation-dependent WRN-RPA interaction promotes recovery of stalled forks at secondary DNA structure |
| title_short | Phosphorylation-dependent WRN-RPA interaction promotes recovery of stalled forks at secondary DNA structure |
| title_sort | phosphorylation dependent wrn rpa interaction promotes recovery of stalled forks at secondary dna structure |
| url | https://doi.org/10.1038/s41467-025-55958-z |
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