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Structure of the replication regulator Sap1 reveals functionally important interfaces
The mechanism by which specific protein-DNA complexes induce programmed replication fork stalling in the eukaryotic genome remains poorly understood. In order to shed light on this process we carried out structural investigations on the essential fission yeast protein Sap1. Sap1 was identified as a...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6053445/ https://www.ncbi.nlm.nih.gov/pubmed/30026545 http://dx.doi.org/10.1038/s41598-018-29198-9 |
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author | Jørgensen, Maria M. Ekundayo, Babatunde Zaratiegui, Mikel Skriver, Karen Thon, Geneviève Schalch, Thomas |
author_facet | Jørgensen, Maria M. Ekundayo, Babatunde Zaratiegui, Mikel Skriver, Karen Thon, Geneviève Schalch, Thomas |
author_sort | Jørgensen, Maria M. |
collection | PubMed |
description | The mechanism by which specific protein-DNA complexes induce programmed replication fork stalling in the eukaryotic genome remains poorly understood. In order to shed light on this process we carried out structural investigations on the essential fission yeast protein Sap1. Sap1 was identified as a protein involved in mating-type switching in Schizosaccharomyces pombe, and has been shown to be involved in programmed replication fork stalling. Interestingly, Sap1 assumes two different DNA binding modes. At the mating-type locus dimers of Sap1 bind the SAS1 sequence in a head-to-head arrangement, while they bind to replication fork blocking sites at rDNA and Tf2 transposons in a head-to-tail mode. In this study, we have solved the crystal structure of the Sap1 DNA binding domain and we observe that Sap1 molecules interact in the crystal using a head-to-tail arrangement that is compatible with DNA binding. We find that Sap1 mutations which alleviate replication-fork blockage at Tf2 transposons in CENP-B mutants map to the head-to-tail interface. Furthermore, several other mutations introduced in this interface are found to be lethal. Our data suggests that essential functions of Sap1 depend on its head-to-tail oligomerization. |
format | Online Article Text |
id | pubmed-6053445 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-60534452018-07-23 Structure of the replication regulator Sap1 reveals functionally important interfaces Jørgensen, Maria M. Ekundayo, Babatunde Zaratiegui, Mikel Skriver, Karen Thon, Geneviève Schalch, Thomas Sci Rep Article The mechanism by which specific protein-DNA complexes induce programmed replication fork stalling in the eukaryotic genome remains poorly understood. In order to shed light on this process we carried out structural investigations on the essential fission yeast protein Sap1. Sap1 was identified as a protein involved in mating-type switching in Schizosaccharomyces pombe, and has been shown to be involved in programmed replication fork stalling. Interestingly, Sap1 assumes two different DNA binding modes. At the mating-type locus dimers of Sap1 bind the SAS1 sequence in a head-to-head arrangement, while they bind to replication fork blocking sites at rDNA and Tf2 transposons in a head-to-tail mode. In this study, we have solved the crystal structure of the Sap1 DNA binding domain and we observe that Sap1 molecules interact in the crystal using a head-to-tail arrangement that is compatible with DNA binding. We find that Sap1 mutations which alleviate replication-fork blockage at Tf2 transposons in CENP-B mutants map to the head-to-tail interface. Furthermore, several other mutations introduced in this interface are found to be lethal. Our data suggests that essential functions of Sap1 depend on its head-to-tail oligomerization. Nature Publishing Group UK 2018-07-19 /pmc/articles/PMC6053445/ /pubmed/30026545 http://dx.doi.org/10.1038/s41598-018-29198-9 Text en © The Author(s) 2018 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/. |
spellingShingle | Article Jørgensen, Maria M. Ekundayo, Babatunde Zaratiegui, Mikel Skriver, Karen Thon, Geneviève Schalch, Thomas Structure of the replication regulator Sap1 reveals functionally important interfaces |
title | Structure of the replication regulator Sap1 reveals functionally important interfaces |
title_full | Structure of the replication regulator Sap1 reveals functionally important interfaces |
title_fullStr | Structure of the replication regulator Sap1 reveals functionally important interfaces |
title_full_unstemmed | Structure of the replication regulator Sap1 reveals functionally important interfaces |
title_short | Structure of the replication regulator Sap1 reveals functionally important interfaces |
title_sort | structure of the replication regulator sap1 reveals functionally important interfaces |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6053445/ https://www.ncbi.nlm.nih.gov/pubmed/30026545 http://dx.doi.org/10.1038/s41598-018-29198-9 |
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