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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...

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Autores principales: 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.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2023
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.
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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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