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DNA-dependent protease activity of human Spartan facilitates replication of DNA–protein crosslink-containing DNA

Mutations in SPARTAN are associated with early onset hepatocellular carcinoma and progeroid features. A regulatory function of Spartan has been implicated in DNA damage tolerance pathways such as translesion synthesis, but the exact function of the protein remained unclear. Here, we reveal the role...

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Autores principales: Mórocz, Mónika, Zsigmond, Eszter, Tóth, Róbert, Enyedi, Márton Zs, Pintér, Lajos, Haracska, Lajos
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5389635/
https://www.ncbi.nlm.nih.gov/pubmed/28053116
http://dx.doi.org/10.1093/nar/gkw1315
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author Mórocz, Mónika
Zsigmond, Eszter
Tóth, Róbert
Enyedi, Márton Zs
Pintér, Lajos
Haracska, Lajos
author_facet Mórocz, Mónika
Zsigmond, Eszter
Tóth, Róbert
Enyedi, Márton Zs
Pintér, Lajos
Haracska, Lajos
author_sort Mórocz, Mónika
collection PubMed
description Mutations in SPARTAN are associated with early onset hepatocellular carcinoma and progeroid features. A regulatory function of Spartan has been implicated in DNA damage tolerance pathways such as translesion synthesis, but the exact function of the protein remained unclear. Here, we reveal the role of human Spartan in facilitating replication of DNA–protein crosslink-containing DNA. We found that purified Spartan has a DNA-dependent protease activity degrading certain proteins bound to DNA. In concert, Spartan is required for direct DPC removal in vivo; we also show that the protease Spartan facilitates repair of formaldehyde-induced DNA–protein crosslinks in later phases of replication using the bromodeoxyuridin (BrdU) comet assay. Moreover, DNA fibre assay indicates that formaldehyde-induced replication stress dramatically decreases the speed of replication fork movement in Spartan-deficient cells, which accumulate in the G2/M cell cycle phase. Finally, epistasis analysis mapped these Spartan functions to the RAD6-RAD18 DNA damage tolerance pathway. Our results reveal that Spartan facilitates replication of DNA–protein crosslink-containing DNA enzymatically, as a protease, which may explain its role in preventing carcinogenesis and aging.
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spelling pubmed-53896352017-04-24 DNA-dependent protease activity of human Spartan facilitates replication of DNA–protein crosslink-containing DNA Mórocz, Mónika Zsigmond, Eszter Tóth, Róbert Enyedi, Márton Zs Pintér, Lajos Haracska, Lajos Nucleic Acids Res Genome Integrity, Repair and Replication Mutations in SPARTAN are associated with early onset hepatocellular carcinoma and progeroid features. A regulatory function of Spartan has been implicated in DNA damage tolerance pathways such as translesion synthesis, but the exact function of the protein remained unclear. Here, we reveal the role of human Spartan in facilitating replication of DNA–protein crosslink-containing DNA. We found that purified Spartan has a DNA-dependent protease activity degrading certain proteins bound to DNA. In concert, Spartan is required for direct DPC removal in vivo; we also show that the protease Spartan facilitates repair of formaldehyde-induced DNA–protein crosslinks in later phases of replication using the bromodeoxyuridin (BrdU) comet assay. Moreover, DNA fibre assay indicates that formaldehyde-induced replication stress dramatically decreases the speed of replication fork movement in Spartan-deficient cells, which accumulate in the G2/M cell cycle phase. Finally, epistasis analysis mapped these Spartan functions to the RAD6-RAD18 DNA damage tolerance pathway. Our results reveal that Spartan facilitates replication of DNA–protein crosslink-containing DNA enzymatically, as a protease, which may explain its role in preventing carcinogenesis and aging. Oxford University Press 2017-04-07 2017-01-02 /pmc/articles/PMC5389635/ /pubmed/28053116 http://dx.doi.org/10.1093/nar/gkw1315 Text en © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Genome Integrity, Repair and Replication
Mórocz, Mónika
Zsigmond, Eszter
Tóth, Róbert
Enyedi, Márton Zs
Pintér, Lajos
Haracska, Lajos
DNA-dependent protease activity of human Spartan facilitates replication of DNA–protein crosslink-containing DNA
title DNA-dependent protease activity of human Spartan facilitates replication of DNA–protein crosslink-containing DNA
title_full DNA-dependent protease activity of human Spartan facilitates replication of DNA–protein crosslink-containing DNA
title_fullStr DNA-dependent protease activity of human Spartan facilitates replication of DNA–protein crosslink-containing DNA
title_full_unstemmed DNA-dependent protease activity of human Spartan facilitates replication of DNA–protein crosslink-containing DNA
title_short DNA-dependent protease activity of human Spartan facilitates replication of DNA–protein crosslink-containing DNA
title_sort dna-dependent protease activity of human spartan facilitates replication of dna–protein crosslink-containing dna
topic Genome Integrity, Repair and Replication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5389635/
https://www.ncbi.nlm.nih.gov/pubmed/28053116
http://dx.doi.org/10.1093/nar/gkw1315
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