Structure of the replication regulator Sap1 reveals functionally important interfaces

Abstract 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 identif...

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Autores principales:	Maria M. Jørgensen, Babatunde Ekundayo, Mikel Zaratiegui, Karen Skriver, Geneviève Thon, Thomas Schalch
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/928c8bb1f12547a5aec8f2641195749b
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spelling	oai:doaj.org-article:928c8bb1f12547a5aec8f2641195749b2021-12-02T15:09:09ZStructure of the replication regulator Sap1 reveals functionally important interfaces10.1038/s41598-018-29198-92045-2322https://doaj.org/article/928c8bb1f12547a5aec8f2641195749b2018-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-29198-9https://doaj.org/toc/2045-2322Abstract 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.Maria M. JørgensenBabatunde EkundayoMikel ZaratieguiKaren SkriverGeneviève ThonThomas SchalchNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-9 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Maria M. Jørgensen Babatunde Ekundayo Mikel Zaratiegui Karen Skriver Geneviève Thon Thomas Schalch Structure of the replication regulator Sap1 reveals functionally important interfaces
description	Abstract 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	article
author	Maria M. Jørgensen Babatunde Ekundayo Mikel Zaratiegui Karen Skriver Geneviève Thon Thomas Schalch
author_facet	Maria M. Jørgensen Babatunde Ekundayo Mikel Zaratiegui Karen Skriver Geneviève Thon Thomas Schalch
author_sort	Maria M. Jørgensen
title	Structure of the replication regulator Sap1 reveals functionally important interfaces
title_short	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_sort	structure of the replication regulator sap1 reveals functionally important interfaces
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/928c8bb1f12547a5aec8f2641195749b
work_keys_str_mv	AT mariamjørgensen structureofthereplicationregulatorsap1revealsfunctionallyimportantinterfaces AT babatundeekundayo structureofthereplicationregulatorsap1revealsfunctionallyimportantinterfaces AT mikelzaratiegui structureofthereplicationregulatorsap1revealsfunctionallyimportantinterfaces AT karenskriver structureofthereplicationregulatorsap1revealsfunctionallyimportantinterfaces AT genevievethon structureofthereplicationregulatorsap1revealsfunctionallyimportantinterfaces AT thomasschalch structureofthereplicationregulatorsap1revealsfunctionallyimportantinterfaces
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Structure of the replication regulator Sap1 reveals functionally important interfaces

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