{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,15]],"date-time":"2026-06-15T13:44:28Z","timestamp":1781531068347,"version":"3.54.5"},"reference-count":32,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2023,3,8]],"date-time":"2023-03-08T00:00:00Z","timestamp":1678233600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["2019R1A2C1008984"],"award-info":[{"award-number":["2019R1A2C1008984"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["R01 ES010546"],"award-info":[{"award-number":["R01 ES010546"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["2019R1A2C1008984"],"award-info":[{"award-number":["2019R1A2C1008984"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 ES010546"],"award-info":[{"award-number":["R01 ES010546"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>DNA polymerase (pol) \u03b7 is responsible for error-free translesion DNA synthesis (TLS) opposite ultraviolet light (UV)-induced cis-syn cyclobutane thymine dimers (CTDs) and cisplatin-induced intrastrand guanine crosslinks. POLH deficiency causes one form of the skin cancer-prone disease xeroderma pigmentosum variant (XPV) and cisplatin sensitivity, but the functional impacts of its germline variants remain unclear. We evaluated the functional properties of eight human POLH germline in silico-predicted deleterious missense variants, using biochemical and cell-based assays. In enzymatic assays, utilizing recombinant pol \u03b7 (residues 1\u2014432) proteins, the C34W, I147N, and R167Q variants showed 4- to 14-fold and 3- to 5-fold decreases in specificity constants (kcat\/Km) for dATP insertion opposite the 3\u2019-T and 5\u2032-T of a CTD, respectively, compared to the wild-type, while the other variants displayed 2- to 4-fold increases. A CRISPR\/Cas9-mediated POLH knockout increased the sensitivity of human embryonic kidney 293 cells to UV and cisplatin, which was fully reversed by ectopic expression of wild-type pol \u03b7, but not by that of an inactive (D115A\/E116A) or either of two XPV-pathogenic (R93P and G263V) mutants. Ectopic expression of the C34W, I147N, and R167Q variants, unlike the other variants, did not rescue the UV- and cisplatin-sensitivity in POLH-knockout cells. Our results indicate that the C34W, I147N, and R167Q variants\u2014substantially reduced in TLS activity\u2014failed to rescue the UV- and cisplatin-sensitive phenotype of POLH-deficient cells, which also raises the possibility that such hypoactive germline POLH variants may increase the individual susceptibility to UV irradiation and cisplatin chemotherapy.<\/jats:p>","DOI":"10.3390\/ijms24065198","type":"journal-article","created":{"date-parts":[[2023,3,9]],"date-time":"2023-03-09T02:33:16Z","timestamp":1678329196000},"page":"5198","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Identification of Three Human POLH Germline Variants Defective in Complementing the UV- and Cisplatin-Sensitivity of POLH-Deficient Cells"],"prefix":"10.3390","volume":"24","author":[{"given":"Mina","family":"Yeom","sequence":"first","affiliation":[{"name":"Department of Pharmacology, Sungkyunkwan University School of Medicine, Suwon 16419, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1302-9462","authenticated-orcid":false,"given":"Jin-Kyung","family":"Hong","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Sungkyunkwan University School of Medicine, Suwon 16419, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0499-4589","authenticated-orcid":false,"given":"Joo-Ho","family":"Shin","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Sungkyunkwan University School of Medicine, Suwon 16419, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0182-2279","authenticated-orcid":false,"given":"Yunjong","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Sungkyunkwan University School of Medicine, Suwon 16419, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7458-3048","authenticated-orcid":false,"given":"Frederick Peter","family":"Guengerich","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN 37232-0146, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeong-Yun","family":"Choi","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Sungkyunkwan University School of Medicine, Suwon 16419, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,3,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"700","DOI":"10.1038\/21447","article-title":"The XPV (xeroderma pigmentosum variant) gene encodes human DNA polymerase \u03b7","volume":"399","author":"Masutani","year":"1999","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"4575","DOI":"10.1021\/bi000130k","article-title":"Efficient translesion replication past oxaliplatin and cisplatin GpG adducts by human DNA polymerase \u03b7","volume":"39","author":"Vaisman","year":"2000","journal-title":"Biochemistry"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"25297","DOI":"10.1074\/jbc.M605699200","article-title":"Biochemical basis of genotoxicity of heterocyclic arylamine food mutagens: Human DNA polymerase \u03b7 selectively produces a two-base deletion in copying the N2-guanyl adduct of 2-amino-3-methylimidazo[4,5-f]quinoline but not the C8 adduct at the NarI G3 site","volume":"281","author":"Choi","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"38244","DOI":"10.1074\/jbc.M608369200","article-title":"Translesion synthesis across O6-alkylguanine DNA adducts by recombinant human DNA polymerases","volume":"281","author":"Choi","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.jmb.2005.06.079","article-title":"Adduct size limits efficient and error-free bypass across bulky N2-guanine DNA lesions by human DNA polymerase \u03b7","volume":"352","author":"Choi","year":"2005","journal-title":"J. Mol. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.jmb.2010.09.015","article-title":"Translesion synthesis across abasic lesions by human B-family and Y-family DNA polymerases \u03b1, \u03b4, \u03b7, \u03b9, \u03ba, and REV1","volume":"404","author":"Choi","year":"2010","journal-title":"J. Mol. Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1044","DOI":"10.1038\/nature09196","article-title":"Structure and mechanism of human DNA polymerase \u03b7","volume":"465","author":"Biertumpfel","year":"2010","journal-title":"Nature"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"7269","DOI":"10.1073\/pnas.1202681109","article-title":"Structural basis of human DNA polymerase \u03b7-mediated chemoresistance to cisplatin","volume":"109","author":"Zhao","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1073\/pnas.022473899","article-title":"Molecular analysis of mutations in DNA polymerase \u03b7 in xeroderma pigmentosum-variant patients","volume":"99","author":"Broughton","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1002\/humu.22462","article-title":"Correlation of phenotype\/genotype in a cohort of 23 xeroderma pigmentosum-variant patients reveals 12 new disease-causing POLH mutations","volume":"35","author":"Opletalova","year":"2014","journal-title":"Hum. Mutat."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"108436","DOI":"10.1016\/j.mrrev.2022.108436","article-title":"Current state of knowledge of human DNA polymerase eta protein structure and disease-causing mutations","volume":"790","author":"Feltes","year":"2022","journal-title":"Mutat. Res. Rev. Mutat. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"979","DOI":"10.2169\/internalmedicine.56.7866","article-title":"Alert Regarding Cisplatin-induced Severe Adverse Events in Cancer Patients with Xeroderma Pigmentosum","volume":"56","author":"Sumiyoshi","year":"2017","journal-title":"Intern. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2473","DOI":"10.1093\/nar\/28.13.2473","article-title":"Complementation of defective translesion synthesis and UV light sensitivity in xeroderma pigmentosum variant cells by human and mouse DNA polymerase \u03b7","volume":"28","author":"Yamada","year":"2000","journal-title":"Nucleic Acids Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"bay119","DOI":"10.1093\/database\/bay119","article-title":"Ensembl variation resources","volume":"2018","author":"Hunt","year":"2018","journal-title":"Database"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"863","DOI":"10.1101\/gr.176601","article-title":"Predicting deleterious amino acid substitutions","volume":"11","author":"Ng","year":"2001","journal-title":"Genome Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1038\/nmeth0410-248","article-title":"A method and server for predicting damaging missense mutations","volume":"7","author":"Adzhubei","year":"2010","journal-title":"Nat. Methods"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1837","DOI":"10.1021\/tx5002755","article-title":"Biochemical analysis of six genetic variants of error-prone human DNA polymerase \u03b9 involved in translesion DNA synthesis","volume":"27","author":"Kim","year":"2014","journal-title":"Chem. Res. Toxicol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2120","DOI":"10.1021\/acs.chemrestox.0c00127","article-title":"Three human pol \u03b9 variants with impaired polymerase activity fail to rescue H2O2 sensitivity in POLI-deficient cells","volume":"33","author":"Yeom","year":"2020","journal-title":"Chem. Res. Toxicol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1741","DOI":"10.1021\/acs.chemrestox.6b00244","article-title":"Six germline genetic variations impair the translesion synthesis activity of human DNA polymerase \u03ba","volume":"29","author":"Kim","year":"2016","journal-title":"Chem. Res. Toxicol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1021\/acs.chemrestox.5b00513","article-title":"Effects of twelve germline missense variations on DNA lesion and G-quadruplex bypass activities of human DNA polymerase REV1","volume":"29","author":"Yeom","year":"2016","journal-title":"Chem. Res. Toxicol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"6469","DOI":"10.1158\/0008-5472.CAN-04-1328","article-title":"The role of DNA polymerase \u03b7 in translesion synthesis past platinum-DNA adducts in human fibroblasts","volume":"64","author":"Bassett","year":"2004","journal-title":"Cancer Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3344","DOI":"10.1128\/MCB.00115-09","article-title":"Human DNA polymerase \u03b7 is required for common fragile site stability during unperturbed DNA replication","volume":"29","author":"Rey","year":"2009","journal-title":"Mol. Cell. Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"D886","DOI":"10.1093\/nar\/gky1016","article-title":"CADD: Predicting the deleteriousness of variants throughout the human genome","volume":"47","author":"Rentzsch","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"E5189","DOI":"10.1073\/pnas.1511585112","article-title":"Comparison of predicted and actual consequences of missense mutations","volume":"112","author":"Miosge","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"572","DOI":"10.1016\/j.ejmg.2017.08.005","article-title":"BRCA1\/2 missense mutations and the value of in-silico analyses","volume":"60","author":"Sadowski","year":"2017","journal-title":"Eur. J. Med. Genet."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3228","DOI":"10.1016\/j.ejca.2009.04.034","article-title":"Variants of the xeroderma pigmentosum variant gene (POLH) are associated with melanoma risk","volume":"45","author":"Guedj","year":"2009","journal-title":"Eur. J. Cancer"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"714","DOI":"10.1016\/j.dnarep.2005.04.002","article-title":"Overproduction of DNA polymerase \u03b7 does not raise the spontaneous mutation rate in diploid human fibroblasts","volume":"4","author":"King","year":"2005","journal-title":"DNA Repair"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"16578","DOI":"10.1073\/pnas.0808589105","article-title":"Human DNA polymerase \u03b7 activity and translocation is regulated by phosphorylation","volume":"105","author":"Chen","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1016\/j.molcel.2009.12.039","article-title":"Regulation of translesion synthesis DNA polymerase \u03b7 by monoubiquitination","volume":"37","author":"Bienko","year":"2010","journal-title":"Mol. Cell"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1016\/S1097-2765(04)00259-X","article-title":"Interaction of human DNA polymerase \u03b7 with monoubiquitinated PCNA: A possible mechanism for the polymerase switch in response to DNA damage","volume":"14","author":"Kannouche","year":"2004","journal-title":"Mol. Cell"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1158\/0008-5472.CAN-05-1862","article-title":"Increased susceptibility to UV-induced skin carcinogenesis in polymerase \u03b7-deficient mice","volume":"66","author":"Lin","year":"2006","journal-title":"Cancer Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1126\/science.1247005","article-title":"Genome-scale CRISPR-Cas9 knockout screening in human cells","volume":"343","author":"Shalem","year":"2014","journal-title":"Science"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/6\/5198\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:51:05Z","timestamp":1760122265000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/6\/5198"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,8]]},"references-count":32,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2023,3]]}},"alternative-id":["ijms24065198"],"URL":"https:\/\/doi.org\/10.3390\/ijms24065198","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,8]]}}}