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By a gel\u2010shift binding assay, we identified an activity in HeLa nuclear extracts that recognizes and binds to oligonucleotides substituted with 2\u2010chloroadenine (ClAde). The activity was specific for ClAde residues because control oligomers did not readily compete out the complex. The binding factor was a monomeric protein that was resistant to inactivation by heating at 45\u00b0C but sensitive to heating at 65\u00b0C, proteinase K treatment, and 5 mM ZnCl<jats:sub>2<\/jats:sub>. This protein, designated ClAde recognition protein (CARP), appeared to be related to a protein that recognized other forms of DNA damage. Gel\u2010shift binding reactions with ultraviolet (UV)\u2010irradiated oligomers revealed a UV\u2010specific protein\/DNA complex that had an electrophoretic mobility similar to that of the CARP\/DNA complex, and CARP binding to ClAde\u2010containing oligomers was readily competed out by UV\u2010irradiated DNA. Moreover, CARP activity was present in extracts prepared from UV\u2010sensitive xeroderma pigmentosum group A cells but not in a subset of cells from group E, suggesting that CARP was similar to a previously described repair associated factor, xeroderma pigmentosum\u2010E binding factor. Our findings support a possible repair process for ClAde residues incorporated into cellular DNA. \u00a9 1995 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/mc.2940130407","type":"journal-article","created":{"date-parts":[[2007,2,21]],"date-time":"2007-02-21T20:56:59Z","timestamp":1172091419000},"page":"245-253","source":"Crossref","is-referenced-by-count":1,"title":["A human factor that recognizes DNA substituted with 2\u2010chloroadenine, an antileukemic purine analog"],"prefix":"10.1002","volume":"13","author":[{"given":"Patricia","family":"Hentosh","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Martin","family":"Tibudan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paul","family":"Grippo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2006,7,20]]},"reference":[{"key":"e_1_2_1_2_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199004193221605"},{"key":"e_1_2_1_3_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V80.9.2203.2203"},{"key":"e_1_2_1_4_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V79.4.882.bloodjournal794882"},{"key":"e_1_2_1_5_2","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.1991.9.3.416"},{"key":"e_1_2_1_6_2","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.1992.10.3.364"},{"key":"e_1_2_1_7_2","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.1992.10.3.371"},{"key":"e_1_2_1_8_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V80.3.587.587"},{"key":"e_1_2_1_9_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4684-1248-2_92"},{"key":"e_1_2_1_10_2","first-page":"6923","article-title":"Mechanisms of inhibition of DNA synthesis by 2\u2010chlorodeoxyadenosine in human lymphoblastic cells","volume":"49","author":"Griffig J","year":"1989","journal-title":"Cancer Res"},{"key":"e_1_2_1_11_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.77.11.6865"},{"key":"e_1_2_1_12_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V62.4.737.737"},{"key":"e_1_2_1_13_2","first-page":"485","article-title":"Interaction of 2\u2010halogenated dATP analogs (F, C, and Br) with human DNA polymerases, DNA primase, and ribonucleotide reductase","volume":"34","author":"Parker WB","year":"1988","journal-title":"Mol Pharmacol"},{"key":"e_1_2_1_14_2","first-page":"915","article-title":"Imbalance of deoxyribonucleoside triphosphates, DNA double\u2010strand breaks, and cell death caused by 2\u2010chlorodeoxyadenosine in mouse FM3A cells","volume":"49","author":"Hirota Y","year":"1989","journal-title":"Cancer Res"},{"key":"e_1_2_1_15_2","doi-asserted-by":"crossref","first-page":"4033","DOI":"10.1016\/S0021-9258(19)39698-X","article-title":"Incorporation of 2\u2010halogeno\u20102\u2010deoxyadenosine 5\u2010triphosphates into DNA during replication by human polymerases \u03b1 and \u03b2","volume":"265","author":"Hentosh P","year":"1990","journal-title":"J Biol Chem"},{"key":"e_1_2_1_16_2","doi-asserted-by":"publisher","DOI":"10.1006\/abbi.1993.1175"},{"key":"e_1_2_1_17_2","first-page":"955","article-title":"Template 2\u2010chloro\u20102\u2032\u2010deoxyadenosine monophosphate inhibits in vitro DNA synthesis","volume":"45","author":"Hentosh P","year":"1994","journal-title":"Mol Pharmacol"},{"key":"e_1_2_1_18_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/19.11.3143"},{"key":"e_1_2_1_19_2","doi-asserted-by":"publisher","DOI":"10.1093\/jnci\/83.8.557"},{"key":"e_1_2_1_20_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00413a022"},{"key":"e_1_2_1_21_2","first-page":"5713","article-title":"Increased DNA repair as a mechanism of acquired resistance to cis\u2010diamminedichloroplatinum (II) in human ovarian cancer cell lines","volume":"48","author":"Masuda H","year":"1988","journal-title":"Cancer Res"},{"key":"e_1_2_1_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.12.9.3689"},{"key":"e_1_2_1_23_2","doi-asserted-by":"publisher","DOI":"10.1096\/fasebj.6.6.1544541"},{"key":"e_1_2_1_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2836(91)90854-Y"},{"key":"e_1_2_1_25_2","doi-asserted-by":"publisher","DOI":"10.1128\/mr.54.1.18-51.1990"},{"key":"e_1_2_1_26_2","doi-asserted-by":"crossref","first-page":"13520","DOI":"10.1016\/S0021-9258(18)42242-9","article-title":"Purification of nuclear proteins that bind to cisplatin\u2010damaged DNA. 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