TopBP1 utilises a bipartite GINS binding mode to support genome replication
Abstract Activation of the replicative Mcm2-7 helicase by loading GINS and Cdc45 is crucial for replication origin firing, and as such for faithful genetic inheritance. Our biochemical and structural studies demonstrate that the helicase activator GINS interacts with TopBP1 through two separate bind...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-02-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-45946-0 |
| _version_ | 1797273968098410496 |
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| author | Matthew Day Bilal Tetik Milena Parlak Yasser Almeida-Hernández Markus Räschle Farnusch Kaschani Heike Siegert Anika Marko Elsa Sanchez-Garcia Markus Kaiser Isabel A. Barker Laurence H. Pearl Antony W. Oliver Dominik Boos |
| author_facet | Matthew Day Bilal Tetik Milena Parlak Yasser Almeida-Hernández Markus Räschle Farnusch Kaschani Heike Siegert Anika Marko Elsa Sanchez-Garcia Markus Kaiser Isabel A. Barker Laurence H. Pearl Antony W. Oliver Dominik Boos |
| author_sort | Matthew Day |
| collection | DOAJ |
| description | Abstract Activation of the replicative Mcm2-7 helicase by loading GINS and Cdc45 is crucial for replication origin firing, and as such for faithful genetic inheritance. Our biochemical and structural studies demonstrate that the helicase activator GINS interacts with TopBP1 through two separate binding surfaces, the first involving a stretch of highly conserved amino acids in the TopBP1-GINI region, the second a surface on TopBP1-BRCT4. The two surfaces bind to opposite ends of the A domain of the GINS subunit Psf1. Mutation analysis reveals that either surface is individually able to support TopBP1-GINS interaction, albeit with reduced affinity. Consistently, either surface is sufficient for replication origin firing in Xenopus egg extracts and becomes essential in the absence of the other. The TopBP1-GINS interaction appears sterically incompatible with simultaneous binding of DNA polymerase epsilon (Polε) to GINS when bound to Mcm2-7-Cdc45, although TopBP1-BRCT4 and the Polε subunit PolE2 show only partial competitivity in binding to Psf1. Our TopBP1-GINS model improves the understanding of the recently characterised metazoan pre-loading complex. It further predicts the coordination of three molecular origin firing processes, DNA polymerase epsilon arrival, TopBP1 ejection and GINS integration into Mcm2-7-Cdc45. |
| first_indexed | 2024-03-07T14:51:45Z |
| format | Article |
| id | doaj.art-91e316aec1f548cab215f4d414dc36db |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-07T14:51:45Z |
| publishDate | 2024-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-91e316aec1f548cab215f4d414dc36db2024-03-05T19:41:15ZengNature PortfolioNature Communications2041-17232024-02-0115112010.1038/s41467-024-45946-0TopBP1 utilises a bipartite GINS binding mode to support genome replicationMatthew Day0Bilal Tetik1Milena Parlak2Yasser Almeida-Hernández3Markus Räschle4Farnusch Kaschani5Heike Siegert6Anika Marko7Elsa Sanchez-Garcia8Markus Kaiser9Isabel A. Barker10Laurence H. Pearl11Antony W. Oliver12Dominik Boos13School of Biological and Behavioural Sciences, Blizard Institute, Queen Mary University of LondonMolecular Genetics II, Center of Medical Biotechnology, University of Duisburg-EssenMolecular Genetics II, Center of Medical Biotechnology, University of Duisburg-EssenComputational Bioengineering, Fakultät Bio- und Chemieingenieurwesen, Technical University DortmundMolecular Genetics, Technical University KaiserslauternAnalytics Core Facility Essen, Center of Medical Biotechnology, University of Duisburg-EssenMolecular Genetics II, Center of Medical Biotechnology, University of Duisburg-EssenMolecular Genetics II, Center of Medical Biotechnology, University of Duisburg-EssenComputational Bioengineering, Fakultät Bio- und Chemieingenieurwesen, Technical University DortmundAnalytics Core Facility Essen, Center of Medical Biotechnology, University of Duisburg-EssenCancer Research UK DNA Repair Enzymes Group, Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, FalmerCancer Research UK DNA Repair Enzymes Group, Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, FalmerCancer Research UK DNA Repair Enzymes Group, Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, FalmerMolecular Genetics II, Center of Medical Biotechnology, University of Duisburg-EssenAbstract Activation of the replicative Mcm2-7 helicase by loading GINS and Cdc45 is crucial for replication origin firing, and as such for faithful genetic inheritance. Our biochemical and structural studies demonstrate that the helicase activator GINS interacts with TopBP1 through two separate binding surfaces, the first involving a stretch of highly conserved amino acids in the TopBP1-GINI region, the second a surface on TopBP1-BRCT4. The two surfaces bind to opposite ends of the A domain of the GINS subunit Psf1. Mutation analysis reveals that either surface is individually able to support TopBP1-GINS interaction, albeit with reduced affinity. Consistently, either surface is sufficient for replication origin firing in Xenopus egg extracts and becomes essential in the absence of the other. The TopBP1-GINS interaction appears sterically incompatible with simultaneous binding of DNA polymerase epsilon (Polε) to GINS when bound to Mcm2-7-Cdc45, although TopBP1-BRCT4 and the Polε subunit PolE2 show only partial competitivity in binding to Psf1. Our TopBP1-GINS model improves the understanding of the recently characterised metazoan pre-loading complex. It further predicts the coordination of three molecular origin firing processes, DNA polymerase epsilon arrival, TopBP1 ejection and GINS integration into Mcm2-7-Cdc45.https://doi.org/10.1038/s41467-024-45946-0 |
| spellingShingle | Matthew Day Bilal Tetik Milena Parlak Yasser Almeida-Hernández Markus Räschle Farnusch Kaschani Heike Siegert Anika Marko Elsa Sanchez-Garcia Markus Kaiser Isabel A. Barker Laurence H. Pearl Antony W. Oliver Dominik Boos TopBP1 utilises a bipartite GINS binding mode to support genome replication Nature Communications |
| title | TopBP1 utilises a bipartite GINS binding mode to support genome replication |
| title_full | TopBP1 utilises a bipartite GINS binding mode to support genome replication |
| title_fullStr | TopBP1 utilises a bipartite GINS binding mode to support genome replication |
| title_full_unstemmed | TopBP1 utilises a bipartite GINS binding mode to support genome replication |
| title_short | TopBP1 utilises a bipartite GINS binding mode to support genome replication |
| title_sort | topbp1 utilises a bipartite gins binding mode to support genome replication |
| url | https://doi.org/10.1038/s41467-024-45946-0 |
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