{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T22:21:43Z","timestamp":1792102903547,"version":"4.4.0"},"reference-count":25,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5425","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1999,7,9]]},"abstract":"<jats:p>\n                    Xeroderma pigmentosum (XP) is an autosomal recessive disease characterized by a high incidence of skin cancers. Yeast\n                    <jats:italic>RAD30<\/jats:italic>\n                    encodes a DNA polymerase involved in the error-free bypass of ultraviolet (UV) damage. Here it is shown that XP variant (XP-V) cell lines harbor nonsense or frameshift mutations in\n                    <jats:italic>hRAD30<\/jats:italic>\n                    , the human counterpart of yeast\n                    <jats:italic>RAD30<\/jats:italic>\n                    . Of the eight mutations identified, seven would result in a severely truncated hRad30 protein. These results indicate that defects in\n                    <jats:italic>hRAD30<\/jats:italic>\n                    cause XP-V, and they suggest that error-free replication of UV lesions by hRad30 plays an important role in minimizing the incidence of sunlight-induced skin cancers.\n                  <\/jats:p>","DOI":"10.1126\/science.285.5425.263","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T05:42:20Z","timestamp":1027748540000},"page":"263-265","source":"Crossref","is-referenced-by-count":637,"title":["<i>hRAD30<\/i>\n                    Mutations in the Variant Form of Xeroderma Pigmentosum"],"prefix":"10.1126","volume":"285","author":[{"given":"Robert E.","family":"Johnson","sequence":"first","affiliation":[{"name":"Sealy Center for Molecular Science, University of Texas Medical Branch at Galveston, 6.104\u00a0Medical Research Building, 11th and Mechanic Streets, Galveston, TX 77555\u20131061, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christine M.","family":"Kondratick","sequence":"additional","affiliation":[{"name":"Sealy Center for Molecular Science, University of Texas Medical Branch at Galveston, 6.104\u00a0Medical Research Building, 11th and Mechanic Streets, Galveston, TX 77555\u20131061, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satya","family":"Prakash","sequence":"additional","affiliation":[{"name":"Sealy Center for Molecular Science, University of Texas Medical Branch at Galveston, 6.104\u00a0Medical Research Building, 11th and Mechanic Streets, Galveston, TX 77555\u20131061, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Louise","family":"Prakash","sequence":"additional","affiliation":[{"name":"Sealy Center for Molecular Science, University of Texas Medical Branch at Galveston, 6.104\u00a0Medical Research Building, 11th and Mechanic Streets, Galveston, TX 77555\u20131061, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/jid.1994.17"},{"key":"e_1_3_1_2_3","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bi.65.070196.000355"},{"key":"e_1_3_1_2_4","unstructured":"; E.\u00a0C.\u00a0Friedberg G.\u00a0C.\u00a0Walker W.\u00a0Siede DNA Repair and Mutagenesis (American Society for Microbiology Washington DC 1995)."},{"key":"e_1_3_1_3_2","doi-asserted-by":"publisher","DOI":"10.1111\/1523-1747.ep12538913"},{"key":"e_1_3_1_3_3","doi-asserted-by":"publisher","DOI":"10.1016\/0921-8777(95)00034-8"},{"key":"e_1_3_1_4_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.72.1.219"},{"key":"e_1_3_1_4_3","doi-asserted-by":"publisher","DOI":"10.1093\/carcin\/1.8.647"},{"key":"e_1_3_1_4_4","doi-asserted-by":"publisher","DOI":"10.1159\/000131646"},{"key":"e_1_3_1_4_5","first-page":"2593","volume":"50","author":"Boyer J. C.","year":"1990","unstructured":"Boyer J. C., et al., Cancer Res. 50, 2593 (1990).","journal-title":"Cancer Res."},{"key":"e_1_3_1_5_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.21.13945"},{"key":"e_1_3_1_5_3","first-page":"2445","volume":"58","author":"Svoboda D. L.","year":"1998","unstructured":"Svoboda D. L., Briley L. P., Vos J.-M. H., Cancer Res. 58, 2445 (1998).","journal-title":"Cancer Res."},{"key":"e_1_3_1_6_2","first-page":"4276","volume":"13","author":"Wang Y.-C.","year":"1993","unstructured":"Wang Y.-C., et al., Mol. Cell. Biol. 13, 4276 (1993);","journal-title":"Mol. Cell. Biol."},{"key":"e_1_3_1_6_3","doi-asserted-by":"publisher","DOI":"10.1111\/1523-1747.ep12371686"},{"key":"e_1_3_1_7_2","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/147.4.1557"},{"key":"e_1_3_1_8_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.274.23.15975"},{"key":"e_1_3_1_9_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.283.5404.1001"},{"key":"e_1_3_1_10_2","unstructured":"The hRAD30 clone was derived from a high-throughput cDNA screening service (Genome Systems St. Louis MO). Oligonucleotides N4920 (5\u2032-CCCGGTACTTGGTGAGGTTAGCTTTCCCACGGG-3\u2032) and N4921 (5\u2032-ATAATTGCAGTGAGTTATGAAGCTCGTGCATTTGGAG-3\u2032) which amplify nucleotides +139 to +280 of the hRAD30 gene were derived from the H96386 cDNA sequence and used to generate a 142-bp PCR fragment. This fragment was used as a probe to screen a human spleen cDNA library. One clone 21749 \u00a0was found to carry a 3.0-kb insert in vector pcDNA2.1 (Invitrogen)."},{"key":"e_1_3_1_11_2","unstructured":"RNA was isolated from various cell lines by use of the Tri-Reagent protocol from Molecular Research Center (Cincinnati OH). hRad30 cDNA was generated from total human RNA by use of the Superscript One Step reverse transcriptase (RT)\u2013PCR system (Gibco-BRL) and amplified in three overlapping fragments. Fragment 1\u00a0was amplified with oligonucleotides RT5\u2032 (5\u2032-GGGAATAAATCTCGCTCGAAACTCACTGGACCGG-3\u2032) and N4920 (9); this fragment encompasses nt \u2212152 to +280 of hRAD30. Fragment 2\u00a0was generated with oligonucleotides N4921 (9) and N4918 (5\u2032-CTTTTGCCTTGATGAGATACGGCAGAAACAACCAGGG-3\u2032); this fragment encompasses nt +139 to +2048 of hRAD30. Fragment 3\u00a0encompasses nt +1738 to +2267 and was amplified with oligonucleotides N4919 (5\u2032-GGGGTGTCGAAGCTAGAAGAATCCTCTAAAGCAACTCC-3\u2032) and RT3\u2032 (5\u2032-TTATTTTTTGTATTAAAAATTTCATAATTCCCTTTCTCAG-3\u2032). Amplified cDNAs from cell lines and clone 21749\u00a0(9) were sequenced with the Thermo Sequenase kit (Amersham Pharmacia Biotech). In addition to the oligonucleotides used for RT-PCR we used the following oligonucleotides as primers to sequence the hRAD30 gene: 5\u2032 Seq (5\u2032-AACTTCTTAGCATCATCTGCCCAC-3\u2032) N5035 (5\u2032-GCCTATCTCGGCAGACTTGTTGCC-3\u2032) N5321 (5\u2032-GAAATGAGAGCAGCCATAGAGAGGG-3\u2032) N5086 (5\u2032-GAGATCCTAGGGATAGAATACATGGG-3\u2032) N5322 (5\u2032-GAGCATCATAGCGGGTAAGGGC-3\u2032) N5087 (5\u2032-CTGGCACCTTTGGCAGAGAACTTGGG-3\u2032) 1565AS (5\u2032-CTCAGTTCCTGTACTTTGACTGG-3\u2032) N5036 (5\u2032-GGGCCAAATCCATCTCTGCAGG-3\u2032) and 3\u2032 Seq (5\u2032-GCAGAAATCCTTTTTGCAGCCCC-3\u2032). Samples were run on 6\u00a0to 8% tris-taurine-EDTA polyacrylamide gels containing 8\u00a0M urea. Gels were fixed in 10% acetic acid 10% methanol solution and dried before autoradiography. Each of the mutations in the XP-V cell lines were independently amplified and sequenced three or four times."},{"key":"e_1_3_1_12_2","unstructured":"Supplemental information is available on Science Online at www.sciencemag.org\/feature\/data\/1041880.shl."},{"key":"e_1_3_1_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/382729a0"},{"key":"e_1_3_1_14_2","doi-asserted-by":"publisher","DOI":"10.1007\/s004380050698"},{"key":"e_1_3_1_15_2","unstructured":"Cell lines were obtained from the Coriell Cell Line Repository (CCR Camden NJ). Lymphoblast cell lines were normal (GM00892B) XPA (GM02250D) XPB (GM02252A) XPC (GM02246C) and XPPHBE (GM02449C). Fibroblast cell lines were XP1CH (GM03055) XP2CH (GM03053) XP115LO (GM02359A) XP30RO (GM03617) XP5MA (GM03379) XP6DU (GM03618) and XP1SF (GM06090)."},{"key":"e_1_3_1_16_2","unstructured":"To quantitate the mutant and wild-type hRad30 transcripts in the XP-V cell lines XPPHBE (GM02449C) and XP1SF (GM06090) we cloned the amplified cDNAs encompassing the respective mutations from each cell line into pUC19-based vectors and sequenced 20\u00a0independent clones from each cell line."},{"key":"e_1_3_1_17_2","unstructured":"C. M.\u00a0Kondratick M.\u00a0T.\u00a0Washington R.\u00a0E.\u00a0Johnson S.\u00a0Prakash L.\u00a0Prakash unpublished observations."},{"key":"e_1_3_1_18_2","unstructured":"Supported by NIH grant GM19261."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.285.5425.263","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,13]],"date-time":"2024-01-13T04:03:42Z","timestamp":1705118622000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.285.5425.263"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1999,7,9]]},"references-count":25,"journal-issue":{"issue":"5425","published-print":{"date-parts":[[1999,7,9]]}},"alternative-id":["10.1126\/science.285.5425.263"],"URL":"https:\/\/doi.org\/10.1126\/science.285.5425.263","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1999,7,9]]}}}