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A disease-associated XPA allele interferes with TFIIH binding and primarily affects transcription-coupled nucleotide excision repair
XPA is a central scaffold protein that coordinates the assembly of repair complexes in the global genome (GG-NER) and transcription-coupled nucleotide excision repair (TC-NER) subpathways. Inactivating mutations in XPA cause xeroderma pigmentosum (XP), which is characterized by extreme UV sensitivit...
Autores principales: | , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
National Academy of Sciences
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10089173/ https://www.ncbi.nlm.nih.gov/pubmed/36893274 http://dx.doi.org/10.1073/pnas.2208860120 |
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author | van den Heuvel, Diana Kim, Mihyun Wondergem, Annelotte P. van der Meer, Paula J. Witkamp, Myrèse Lambregtse, Ferdy Kim, Hyun-Suk Kan, Folkert Apelt, Katja Kragten, Angela González-Prieto, Román Vertegaal, Alfred C. O. Yeo, Jung-Eun Kim, Byung-Gyu van Doorn, Remco Schärer, Orlando D. Luijsterburg, Martijn S. |
author_facet | van den Heuvel, Diana Kim, Mihyun Wondergem, Annelotte P. van der Meer, Paula J. Witkamp, Myrèse Lambregtse, Ferdy Kim, Hyun-Suk Kan, Folkert Apelt, Katja Kragten, Angela González-Prieto, Román Vertegaal, Alfred C. O. Yeo, Jung-Eun Kim, Byung-Gyu van Doorn, Remco Schärer, Orlando D. Luijsterburg, Martijn S. |
author_sort | van den Heuvel, Diana |
collection | PubMed |
description | XPA is a central scaffold protein that coordinates the assembly of repair complexes in the global genome (GG-NER) and transcription-coupled nucleotide excision repair (TC-NER) subpathways. Inactivating mutations in XPA cause xeroderma pigmentosum (XP), which is characterized by extreme UV sensitivity and a highly elevated skin cancer risk. Here, we describe two Dutch siblings in their late forties carrying a homozygous H244R substitution in the C-terminus of XPA. They present with mild cutaneous manifestations of XP without skin cancer but suffer from marked neurological features, including cerebellar ataxia. We show that the mutant XPA protein has a severely weakened interaction with the transcription factor IIH (TFIIH) complex leading to an impaired association of the mutant XPA and the downstream endonuclease ERCC1-XPF with NER complexes. Despite these defects, the patient-derived fibroblasts and reconstituted knockout cells carrying the XPA-H244R substitution show intermediate UV sensitivity and considerable levels of residual GG-NER (~50%), in line with the intrinsic properties and activities of the purified protein. By contrast, XPA-H244R cells are exquisitely sensitive to transcription-blocking DNA damage, show no detectable recovery of transcription after UV irradiation, and display a severe deficiency in TC-NER-associated unscheduled DNA synthesis. Our characterization of a new case of XPA deficiency that interferes with TFIIH binding and primarily affects the transcription-coupled subpathway of nucleotide excision repair, provides an explanation of the dominant neurological features in these patients, and reveals a specific role for the C-terminus of XPA in TC-NER. |
format | Online Article Text |
id | pubmed-10089173 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | National Academy of Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-100891732023-04-12 A disease-associated XPA allele interferes with TFIIH binding and primarily affects transcription-coupled nucleotide excision repair van den Heuvel, Diana Kim, Mihyun Wondergem, Annelotte P. van der Meer, Paula J. Witkamp, Myrèse Lambregtse, Ferdy Kim, Hyun-Suk Kan, Folkert Apelt, Katja Kragten, Angela González-Prieto, Román Vertegaal, Alfred C. O. Yeo, Jung-Eun Kim, Byung-Gyu van Doorn, Remco Schärer, Orlando D. Luijsterburg, Martijn S. Proc Natl Acad Sci U S A Biological Sciences XPA is a central scaffold protein that coordinates the assembly of repair complexes in the global genome (GG-NER) and transcription-coupled nucleotide excision repair (TC-NER) subpathways. Inactivating mutations in XPA cause xeroderma pigmentosum (XP), which is characterized by extreme UV sensitivity and a highly elevated skin cancer risk. Here, we describe two Dutch siblings in their late forties carrying a homozygous H244R substitution in the C-terminus of XPA. They present with mild cutaneous manifestations of XP without skin cancer but suffer from marked neurological features, including cerebellar ataxia. We show that the mutant XPA protein has a severely weakened interaction with the transcription factor IIH (TFIIH) complex leading to an impaired association of the mutant XPA and the downstream endonuclease ERCC1-XPF with NER complexes. Despite these defects, the patient-derived fibroblasts and reconstituted knockout cells carrying the XPA-H244R substitution show intermediate UV sensitivity and considerable levels of residual GG-NER (~50%), in line with the intrinsic properties and activities of the purified protein. By contrast, XPA-H244R cells are exquisitely sensitive to transcription-blocking DNA damage, show no detectable recovery of transcription after UV irradiation, and display a severe deficiency in TC-NER-associated unscheduled DNA synthesis. Our characterization of a new case of XPA deficiency that interferes with TFIIH binding and primarily affects the transcription-coupled subpathway of nucleotide excision repair, provides an explanation of the dominant neurological features in these patients, and reveals a specific role for the C-terminus of XPA in TC-NER. National Academy of Sciences 2023-03-09 2023-03-14 /pmc/articles/PMC10089173/ /pubmed/36893274 http://dx.doi.org/10.1073/pnas.2208860120 Text en Copyright © 2023 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) . |
spellingShingle | Biological Sciences van den Heuvel, Diana Kim, Mihyun Wondergem, Annelotte P. van der Meer, Paula J. Witkamp, Myrèse Lambregtse, Ferdy Kim, Hyun-Suk Kan, Folkert Apelt, Katja Kragten, Angela González-Prieto, Román Vertegaal, Alfred C. O. Yeo, Jung-Eun Kim, Byung-Gyu van Doorn, Remco Schärer, Orlando D. Luijsterburg, Martijn S. A disease-associated XPA allele interferes with TFIIH binding and primarily affects transcription-coupled nucleotide excision repair |
title | A disease-associated XPA allele interferes with TFIIH binding and primarily affects transcription-coupled nucleotide excision repair |
title_full | A disease-associated XPA allele interferes with TFIIH binding and primarily affects transcription-coupled nucleotide excision repair |
title_fullStr | A disease-associated XPA allele interferes with TFIIH binding and primarily affects transcription-coupled nucleotide excision repair |
title_full_unstemmed | A disease-associated XPA allele interferes with TFIIH binding and primarily affects transcription-coupled nucleotide excision repair |
title_short | A disease-associated XPA allele interferes with TFIIH binding and primarily affects transcription-coupled nucleotide excision repair |
title_sort | disease-associated xpa allele interferes with tfiih binding and primarily affects transcription-coupled nucleotide excision repair |
topic | Biological Sciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10089173/ https://www.ncbi.nlm.nih.gov/pubmed/36893274 http://dx.doi.org/10.1073/pnas.2208860120 |
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