{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T13:52:59Z","timestamp":1791121979762,"version":"4.1.0"},"reference-count":57,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5416","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1999,5,7]]},"abstract":"<jats:p>To study the nuclear organization and dynamics of nucleotide excision repair (NER), the endonuclease ERCC1\/XPF (for excision repair cross complementation group 1\/xeroderma pigmentosum group F) was tagged with green fluorescent protein and its mobility was monitored in living Chinese hamster ovary cells. In the absence of DNA damage, the complex moved freely through the nucleus, with a diffusion coefficient (15 \u00b1 5 square micrometers per second) consistent with its molecular size. Ultraviolet light\u2013induced DNA damage caused a transient dose-dependent immobilization of ERCC1\/XPF, likely due to engagement of the complex in a single repair event. After 4 minutes, the complex regained mobility. These results suggest (i) that NER operates by assembly of individual NER factors at sites of DNA damage rather than by preassembly of holocomplexes and (ii) that ERCC1\/XPF participates in repair of DNA damage in a distributive fashion rather than by processive scanning of large genome segments.<\/jats:p>","DOI":"10.1126\/science.284.5416.958","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:50:55Z","timestamp":1027763455000},"page":"958-961","source":"Crossref","is-referenced-by-count":284,"title":["Action of DNA Repair Endonuclease ERCC1\/XPF in Living Cells"],"prefix":"10.1126","volume":"284","author":[{"given":"Adriaan B.","family":"Houtsmuller","sequence":"first","affiliation":[{"name":"Department of Pathology (Josephine Nefkens Institute) and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suzanne","family":"Rademakers","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Genetics (Medical Genetics Center, CBG), Erasmus University, Post Office Box 1738, 3000 DR Rotterdam, Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alex L.","family":"Nigg","sequence":"additional","affiliation":[{"name":"Department of Pathology (Josephine Nefkens Institute) and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deborah","family":"Hoogstraten","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Genetics (Medical Genetics Center, CBG), Erasmus University, Post Office Box 1738, 3000 DR Rotterdam, Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jan H.","family":"J.","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"family":"Hoeijmakers","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Genetics (Medical Genetics Center, CBG), Erasmus University, Post Office Box 1738, 3000 DR Rotterdam, Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wim","family":"Vermeulen","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Genetics (Medical Genetics Center, CBG), Erasmus University, Post Office Box 1738, 3000 DR Rotterdam, Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_2_2_2","unstructured":"E. 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The operational diffusion coefficient of EGFP was in the same range as previously reported values measured with conventional spot-FRAP (18) ["},{"key":"e_1_3_2_20_6","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-3495(97)78835-0"},{"key":"e_1_3_2_21_2","unstructured":"For a comparison between MW and D see"},{"key":"e_1_3_2_21_3","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.138.1.131"},{"key":"e_1_3_2_22_2","unstructured":"XPA and XPB were tagged with EGFP respectively at the NH 2 - and COOH-termini (S. Rademakers D. Hoogstraten W. Vermeulen unpublished data) and stably expressed in SV40-transformed XP-A and XP-B cells."},{"key":"e_1_3_2_23_2","unstructured":"RAD52 is involved in homologous recombination repair of x ray\u2013induced DNA damage ["},{"key":"e_1_3_2_23_3","doi-asserted-by":"crossref","first-page":"6423","DOI":"10.1128\/MCB.18.11.6423","volume":"18","author":"Rijkers T.","year":"1998","unstructured":"Rijkers T., et al., Mol. Cell. Biol. 18, 6423 (1998)].","journal-title":"Mol. Cell. Biol."},{"key":"e_1_3_2_24_2","unstructured":"The androgen receptor is involved in steroid hormone\u2013dependent transcription regulation."},{"key":"e_1_3_2_25_2","unstructured":"Global genome repair in rodent cells removes predominantly 6-4PPs ["},{"key":"e_1_3_2_25_3","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/S0165-7992(85)80018-X","volume":"143","author":"Mitchel D. L.","year":"1985","unstructured":"Mitchel D. L., Haipek C. A., Clarkson J. M, Mutat. Res. 143, 109 (1985);","journal-title":"Mutat. Res."},{"key":"e_1_3_2_25_4","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1111\/j.1751-1097.1989.tb05578.x","volume":"49","author":"Mitchell D. L.","year":"1989","unstructured":"Mitchell D. L., Nairn R. S., Photochem. Photobiol. 49, 805 (1989)].","journal-title":"Photochem. Photobiol."},{"key":"e_1_3_2_26_2","unstructured":"Given that a nucleus contains \u223c30 000 ERCC1-GFP\/XPF complexes (J. H. J. Hoeijmakers unpublished results) of which one-sixth (averaged over the whole repair period; Fig. 4C) is involved in NER we estimate that damage is repaired at a rate of 1250 lesions per minute (1\/6 \u00d7 30 000\/4 min). Thus repair of 300 000 6-4PP lesions (16) induced by UV light (8 J\/m 2 ) would require \u223c4 hours (Fig. 4C)."},{"key":"e_1_3_2_27_2","doi-asserted-by":"crossref","first-page":"1336","DOI":"10.1016\/S0021-9258(19)85091-3","volume":"264","author":"Oh E. Y.","year":"1989","unstructured":"Oh E. Y., Grossman L., J. Biol. Chem. 264, 1336 (1989).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_2_28_2","doi-asserted-by":"crossref","first-page":"2877","DOI":"10.1093\/nar\/24.15.2877","volume":"24","author":"Koehler D. R.","year":"1996","unstructured":"Koehler D. R., Hanawalt P. C., Nucleic Acids Res. 24, 2877 (1996).","journal-title":"Nucleic Acids Res."},{"key":"e_1_3_2_29_2","unstructured":"Immunoblot analysis of total cell extracts (10 \u03bcg) and immunoprecipitation of cell lysates were performed as described (8). A standard colony-survival procedure for assessment of UV sensitivity (8) was applied to CHO-9 (parental repair-competent) 43-3B (ERCC1\/repair-deficient) and 43-3B (ERCC1-GFP transfected) cells. At 5 days after UV irradiation (Philips TUV-lamp) the cell survival rate [(number of surviving treated colonies)\/(number of surviving untreated colonies)] was plotted against UV dose."},{"key":"e_1_3_2_30_2","unstructured":"We thank P. Berkhout for statistical advice; G. van Cappellen for technical advice; J. Essers and P. Doesburg for cell lines expressing respectively RAD52-GFP and AR-GFP; R. van Geloof for software; and R. van Driel L. H. F. Mullenders A. A. van Zeeland and J. Trapman for helpful suggestions. Supported by the Dutch Cancer Society (KWF) the Dutch Scientific Organization (NWO) and by the Louis Jeantet Foundation."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.284.5416.958","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,13]],"date-time":"2024-01-13T09:29:20Z","timestamp":1705138160000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.284.5416.958"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1999,5,7]]},"references-count":57,"journal-issue":{"issue":"5416","published-print":{"date-parts":[[1999,5,7]]}},"alternative-id":["10.1126\/science.284.5416.958"],"URL":"https:\/\/doi.org\/10.1126\/science.284.5416.958","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1999,5,7]]}}}