{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T05:36:31Z","timestamp":1791178591434,"version":"4.1.0"},"reference-count":45,"publisher":"The Company of Biologists","issue":"Supplement_6","license":[{"start":{"date-parts":[[1987,1,1]],"date-time":"1987-01-01T00:00:00Z","timestamp":536457600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/www.biologists.com\/user-licence-1-1\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[1987,1,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n               <jats:p>To extend our knowledge of the excision repair system in mammalian cells we have focussed on the isolation of genes and proteins involved in this process. For the purification and characterization of human repair proteins the microneedle injection assay technique is utilized. This system is based on the transient correction of the excision repair defect of xeroderma pigmentosum (XP) fibroblasts (scored as increase of ultraviolet (u.v.)-induced unscheduled DNA synthesis (UDS)) upon microinjection of crude extracts from complementing XP or normal cells. Specific correction is observed in fibroblasts of all (9) excision-deficient XP complementation groups. The XP-A and G correcting factors were found to be proteins and several purification steps (including (NH4)2SO4 fractionation, chromatography of phosphocellulose, heparin and u.v.- irradiated DNA-cellulose) have been worked out for the XP-A correcting protein.<\/jats:p>\n               <jats:p>The microinjection system was also used for the introduction of (partially) purified repair enzymes of lower organisms. Micrococcus luteus endonuclease and bacteriophage T4 endonuclease V were able to correct all XP complementation groups tested, in marked contrast to the more sophisticated Escherichia coli uvrABC complex injected with uvrD. Photoreversal of dimers could be registered after introduction of the yeast photoreactivating enzyme in repair-competent, XP- variant, XP-C and XP-I fibroblasts (monitored as decrease of (residual) UDS). Remarkably, no effect was noticed in XP-A, D, E and H, suggesting that something prevents dimers in these cells from being monomerized by the injected enzyme.<\/jats:p>\n               <jats:p>Using DNA-mediated gene transfer we have cloned a human gene (designated ERCC-1) that compensates for the excision defect of the u.v. and mitomycin C-sensitive Chinese hamster ovary cell (CHO) mutant 43-3B (complementation group 2). Characterization of this gene and its cDNA revealed the following features:<\/jats:p>\n               <jats:p>(1) ERCC-1 corrects the full spectrum of repair deficiencies in mutants of complementation group 2. No correction is observed in mutants of the other CHO complementation groups.<\/jats:p>\n               <jats:p>(2) The ERCC-l gene has a size of 15 \u00d7101 base-pairs (bp) and consists of 10 exons, one of which appears to be differentially spliced.<\/jats:p>\n               <jats:p>(3) It encodes two largely identical mRNAs, which differ in the presence or absence of a 72bp coding exon, situated in the 3 \u2032 half of the mRNA. Only the cDNA of the large transcript is able to confer repair proficiency to 43-3B cells. No effect of u.v. treatment is found at the level of ERCC-1 transcription in HeLa cells.<\/jats:p>\n               <jats:p>(4) Sequence analysis of full-length cDNA copies of the two ERCC-1 mRNAs revealed open reading frames for proteins of 297 and 273 amino acids, respectively. Significant amino acid sequence homology was found between portions of the putative ERCC-1 product and the protein encoded by the yeast excision-repair gene RAD10. Regional homology was also discovered between a part of ERCC-1 and uvrA. On the basis of homology with functional protein domains a tentative nuclear location signal, DNA binding domain and ADP-ribosylation site could be identified in the ERCC-1 aa sequence.<\/jats:p>","DOI":"10.1242\/jcs.1984.supplement_6.7","type":"journal-article","created":{"date-parts":[[2013,5,23]],"date-time":"2013-05-23T07:59:48Z","timestamp":1369295988000},"page":"111-125","source":"Crossref","is-referenced-by-count":18,"title":["Characterization of Genes and Proteins Involved in Excision Repair of Human Cells"],"prefix":"10.1242","volume":"1987","author":[{"given":"Jan H. J.","family":"Hoeijmakers","sequence":"first","affiliation":[{"name":"Department of Cell Biology and Genetics, Erasmus University, PO Box 1738, 3000 DR Rotterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","published-online":{"date-parts":[[1987,1,1]]},"reference":[{"key":"2024071019364456400_JOCES_1987_S6_111C1","first-page":"2735","article-title":"Primary structure of the RAD52 gene in Saccharomyces cerevisiae.","volume":"4","author":"Adzuma","year":"1984","journal-title":"Molec. cell. Biol."},{"key":"2024071019364456400_JOCES_1987_S6_111C2","doi-asserted-by":"crossref","first-page":"652","DOI":"10.1038\/218652a0","article-title":"Defective repair replication of DNA in xeroderma pigmentosum","volume":"218","author":"Cleaver","year":"1968","journal-title":"Nature, Lond."},{"key":"2024071019364456400_JOCES_1987_S6_111C3","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/0027-5107(85)90106-X","article-title":"Microinjection of Micrococcus luteus UV-endonuclease restores UV-induced unscheduled DNA synthesis in cells of 9 xeroderma pigmentosum complementation groups","volume":"150","author":"de Jonge","year":"1985","journal-title":"Mutat. Res."},{"key":"2024071019364456400_JOCES_1987_S6_111C4","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1002\/j.1460-2075.1983.tb01477.x","article-title":"Microinjection of human cell extracts corrects xeroderma pigmentosum defect","volume":"2","author":"de Jonge","year":"1983","journal-title":"EMBOJ."},{"key":"2024071019364456400_JOCES_1987_S6_111C5","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1038\/newbio238080a0","article-title":"Genetic heterogeneity of xeroderma pigmentosum demonstrated by somatic cell hybridization","volume":"238","author":"de Weerd-Kastelein","year":"1972","journal-title":"Nature, new Biol."},{"key":"2024071019364456400_JOCES_1987_S6_111C6","first-page":"217","article-title":"A ninth complementation group in xeroderma pigmentosum, XP-I","volume":"145","author":"Fischer","year":"1985","journal-title":"Mutat. Res."},{"key":"2024071019364456400_JOCES_1987_S6_111C7","volume-title":"DNARepair. San Francisco: Freeman & Company","author":"Friedberg","year":"1985"},{"key":"2024071019364456400_JOCES_1987_S6_111C8","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1016\/0092-8674(82)90161-1","article-title":"Differences in patterns of complementation of the more common groups of xeroderma pigmentosum: possible implications","volume":"29","author":"Giannelli","year":"1982","journal-title":"Cell"},{"key":"2024071019364456400_JOCES_1987_S6_111C9","doi-asserted-by":"crossref","first-page":"3465","DOI":"10.1016\/S0021-9258(18)33472-0","article-title":"Enzymatic repair of pyrimidine dimercontaining DNA: a 5\u2019-dimer DNA glycosylase-3\u2019 apyrimidinic endonuclease mechanism from Micrococcus luteus.","volume":"257","author":"GRAFSTROM","year":"1982","journal-title":"J. biol. Chem."},{"key":"2024071019364456400_JOCES_1987_S6_111C10","first-page":"634","volume":"285","author":"Haseltine","year":"1980","journal-title":"Cleavage of pyrimidine dimers in specific DNA sequences by a pyrimidine dimer DNA-glycosylase of M. luteus. Nature, Lond."},{"key":"2024071019364456400_JOCES_1987_S6_111C11","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1016\/0027-5107(81)90110-X","article-title":"Repair of ultraviolet radiation damage in xeroderma pigmentosum cells belonging to complementation group F","volume":"80","author":"Hayakawa","year":"1981","journal-title":"Mutat. Res."},{"key":"2024071019364456400_JOCES_1987_S6_111C12","doi-asserted-by":"crossref","DOI":"10.1016\/0014-4827(87)90230-8","article-title":"Differences between rodent and human cell lines in the amount of integrated DNA after transfection","volume-title":"Expl Cell Res. (in press)","author":"Hoeijmakers","year":"1987"},{"key":"2024071019364456400_JOCES_1987_S6_111C13","article-title":"Identification of DNA-repair genes in the human genome","volume-title":"Cold Spring Harbor. Symp. quant. Biol, (in press)","author":"Hoeijmakers","year":"1987"},{"key":"2024071019364456400_JOCES_1987_S6_111C14","doi-asserted-by":"crossref","first-page":"4895","DOI":"10.1016\/S0021-9258(19)89189-5","article-title":"Sequence of Escherichia coliuvr A gene and protein reveal two potential ATP binding sites","volume":"261","author":"Husain","year":"1986","journal-title":"J. biol. 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U.S A."},{"key":"2024071019364456400_JOCES_1987_S6_111C17","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/0027-5107(85)90101-0","article-title":"Use of recombinant DNA techniques in cloning DNA repair genes and in the study of mutagenesis in human cells","volume":"150","author":"Lehmann","year":"1985","journal-title":"Mutat. Res."},{"key":"2024071019364456400_JOCES_1987_S6_111C18","first-page":"1152","article-title":"DNA-mediated cotransfer of excision repair capacity and drug resistance into Chinese hamster ovary cell line UV-135","volume":"4","author":"MacInnes","year":"1984","journal-title":"Molec. cell. Biol."},{"key":"2024071019364456400_JOCES_1987_S6_111C19","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1016\/S0076-6879(80)65059-9","article-title":"Sequence end-labeled DNA with base-specific chemical cleavages","volume":"65","author":"Maxam","year":"1980","journal-title":"Meth. Enzym."},{"key":"2024071019364456400_JOCES_1987_S6_111C20","first-page":"17","article-title":"RAD3 gene of Saccharomyces cerevisiae: nucleotide sequence of wild type and mutant alleles, transcript mapping, and aspects of gene regulation","volume":"5","author":"Naumovski","year":"1985","journal-title":"Molec. cell. Biol."},{"key":"2024071019364456400_JOCES_1987_S6_111C21","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/0378-1119(85)90177-5","article-title":"The RAD2 gene of Saccharomyces cerevisiae-. nucleotide sequence and transcript mapping","volume":"36","author":"Nicolet","year":"1985","journal-title":"Gene"},{"key":"2024071019364456400_JOCES_1987_S6_111C22","first-page":"280","article-title":"A cDNA cloning vector that permits expression of cDNA inserts in mammalian cell","volume":"3","author":"Okayama","year":"1983","journal-title":"Molec. cell. Biol."},{"key":"2024071019364456400_JOCES_1987_S6_111C23","first-page":"1497","article-title":"RAD7 gene of Saccharomyces cerevisae-. Transcripts, nucleotide sequence analysis, and functional relationship between the RAD7 and RAD23 gene products","volume":"6","author":"Perozzi","year":"1986","journal-title":"Molec. cell. Biol."},{"key":"2024071019364456400_JOCES_1987_S6_111C24","doi-asserted-by":"crossref","first-page":"3549","DOI":"10.1002\/j.1460-2075.1985.tb04115.x","article-title":"Nucleotide sequence of the RADIO gene of Saccharomyces cerevisiae.","volume":"4","author":"Reynolds","year":"1985","journal-title":"EMBOy."},{"key":"2024071019364456400_JOCES_1987_S6_111C25","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1073\/pnas.82.1.168","article-title":"RAD6 gene of Saccharomyces cerevisiaeencodes a protein containing a tract of 13 consecutive aspartates","volume":"82","author":"Reynolds","year":"1985","journal-title":"Proc. natn. Acad. Sci. U.SA."},{"key":"2024071019364456400_JOCES_1987_S6_111C26","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1038\/311390a0","article-title":"Isolation of UV-resistant revertants from a xeroderma pigmentosum complementation group A cell line","volume":"311","author":"Royer-Pokora","year":"1984","journal-title":"Nature, Lond."},{"key":"2024071019364456400_JOCES_1987_S6_111C27","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1038\/306206a0","article-title":"Molecular identification of a human repair gene following DNA-mediated gene transfer","volume":"306","author":"Rubin","year":"1983","journal-title":"Nature, Lond."},{"key":"2024071019364456400_JOCES_1987_S6_111C28","first-page":"353","volume-title":"In Atlas of Protein Sequence and Structure","author":"Schwartz","year":"1978"},{"key":"2024071019364456400_JOCES_1987_S6_111C29","doi-asserted-by":"crossref","first-page":"2958","DOI":"10.1073\/pnas.74.7.2958","article-title":"Specific action of T4 endonuclease V on damaged DNA in xeroderma pigmentosum cells in vivo.","volume":"74","author":"Tanaka","year":"1977","journal-title":"Proc. natn. Acad. Sci. U.SA."},{"key":"2024071019364456400_JOCES_1987_S6_111C30","doi-asserted-by":"crossref","first-page":"4071","DOI":"10.1073\/pnas.72.10.4071","article-title":"Restoration of ultraviolet-induced unscheduled DNA synthesis of xeroderma pigmentosum cells by the concomittant treatment with bacteriophage T4 endonuclease V and HVJ (Sendai virus)","volume":"72","author":"Tanaka","year":"1975","journal-title":"Proc. natn. Acad. Sci. U.SA."},{"key":"2024071019364456400_JOCES_1987_S6_111C31","first-page":"315","article-title":"The intracellular signal for induction of resistance to alkylating agents","volume":"48","author":"Teo","year":"1986","journal-title":"E. coli. Cell"},{"key":"2024071019364456400_JOCES_1987_S6_111C32","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1007\/BF01543017","article-title":"Hypersensitivity to mutation and sister-chromatid exchange induction in CHO cell mutants defective in incising DNA containing UV-lesions","volume":"8","author":"Thompson","year":"1982","journal-title":"Somat. Cell Genet."},{"key":"2024071019364456400_JOCES_1987_S6_111C33","first-page":"881","article-title":"DNA-mediated transfer of a human DNA repair gene that controls sister chromatid exchange","volume":"5","author":"Thompson","year":"1985","journal-title":"Molec. cell. Biol."},{"key":"2024071019364456400_JOCES_1987_S6_111C34","doi-asserted-by":"crossref","first-page":"3734","DOI":"10.1073\/pnas.78.6.3734","article-title":"Genetic diversity of UV-sensitive DNA-repair mutants of Chinese hamster ovary cells","volume":"78","author":"Thompson","year":"1981","journal-title":"Proc, natn. Acad. Sci. U.SA."},{"key":"2024071019364456400_JOCES_1987_S6_111C35","first-page":"125","volume-title":"Cellular Responses to DNA Damage","author":"Thompson","year":"1983"},{"key":"2024071019364456400_JOCES_1987_S6_111C36","first-page":"913","volume":"44","author":"Van Duin","year":"1986","journal-title":"Molecular characterization of the human excision repair gene ERCC-1: cDNA cloning and amino acid homology with the yeast DNA repair gene RADIO. Cell"},{"key":"2024071019364456400_JOCES_1987_S6_111C37","first-page":"199","article-title":"Transient correction of excision repair defects in fibroblasts of 9 xeroderma pigmentosum complementation groups by microinjection of crude human cell extracts","volume":"165","author":"Vermeulen","year":"1986","journal-title":"Mutat. 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