{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T03:21:10Z","timestamp":1772162470731,"version":"3.50.1"},"reference-count":30,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5543","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[2001,10,26]]},"abstract":"<jats:p>\n                    In response to DNA damage, eukaryotic cells activate checkpoint pathways that arrest cell cycle progression and induce the expression of genes required for DNA repair. In budding yeast, the homothallic switching (HO) endonuclease creates a site-specific double-strand break at the mating type (\n                    <jats:italic>MAT<\/jats:italic>\n                    ) locus. Continuous\n                    <jats:italic>HO<\/jats:italic>\n                    expression results in the phosphorylation of Rad53, which is dependent on products of the ataxia telangiectasia mutated\u2013related\n                    <jats:italic>MEC1<\/jats:italic>\n                    gene and other checkpoint genes, including\n                    <jats:italic>DDC1<\/jats:italic>\n                    ,\n                    <jats:italic>RAD9<\/jats:italic>\n                    , and\n                    <jats:italic>RAD24<\/jats:italic>\n                    . Chromatin immunoprecipitation experiments revealed that the Ddc1 protein associates with a region near the\n                    <jats:italic>MAT<\/jats:italic>\n                    locus after\n                    <jats:italic>HO<\/jats:italic>\n                    expression. Ddc1 association required Rad24 but not Mec1 or Rad9. Mec1 also associated with a region near the cleavage site after\n                    <jats:italic>HO<\/jats:italic>\n                    expression, but this association is independent of Ddc1, Rad9, and Rad24. Thus, Mec1 and Ddc1 are recruited independently to sites of DNA damage, suggesting the existence of two separate mechanisms involved in recognition of DNA damage.\n                  <\/jats:p>","DOI":"10.1126\/science.1063827","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T05:54:32Z","timestamp":1027749272000},"page":"867-870","source":"Crossref","is-referenced-by-count":204,"title":["Recruitment of Mec1 and Ddc1 Checkpoint Proteins to Double-Strand Breaks Through Distinct Mechanisms"],"prefix":"10.1126","volume":"294","author":[{"given":"Tae","family":"Kondo","sequence":"first","affiliation":[{"name":"Division of Biological Science, Graduate School of Science, Nagoya University,"}]},{"given":"Tatsushi","family":"Wakayama","sequence":"additional","affiliation":[{"name":"Division of Biological Science, Graduate School of Science, Nagoya University,"}]},{"given":"Takahiro","family":"Naiki","sequence":"additional","affiliation":[{"name":"Division of Biological Science, Graduate School of Science, Nagoya University,"}]},{"given":"Kunihiro","family":"Matsumoto","sequence":"additional","affiliation":[{"name":"Division of Biological Science, Graduate School of Science, Nagoya University,"},{"name":"CREST, Japan Science and Technology Corporation, Chikusa-ku, Nagoya 464-0814, Japan."}]},{"given":"Katsunori","family":"Sugimoto","sequence":"additional","affiliation":[{"name":"Division of Biological Science, Graduate School of Science, Nagoya University,"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/35044005"},{"key":"e_1_3_1_3_2","doi-asserted-by":"crossref","first-page":"5525","DOI":"10.1093\/emboj\/17.19.5525","volume":"17","author":"Longhese M. P.","year":"1998","unstructured":"Longhese M. P., Foiani M., Muzi-Falconi M., Lucchini G., Plevani P., EMBO J. 17, 5525 (1998).","journal-title":"EMBO J."},{"key":"e_1_3_1_4_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.274.5293.1664"},{"key":"e_1_3_1_5_2","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/S0027-5107(00)00040-3","volume":"451","author":"Haber J. E.","year":"2000","unstructured":"Haber J. E., Mutat. Res. 451, 53 (2000).","journal-title":"Mutat. Res."},{"key":"e_1_3_1_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(93)90493-A"},{"key":"e_1_3_1_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)80375-X"},{"key":"e_1_3_1_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)81482-8"},{"key":"e_1_3_1_9_2","doi-asserted-by":"crossref","first-page":"1891","DOI":"10.1128\/MCB.18.4.1891","volume":"18","author":"Lewis L. K.","year":"1998","unstructured":"Lewis L. K., Kirchner J. M., Resnick M. A., Mol. Cell. Biol. 18, 1891 (1998).","journal-title":"Mol. Cell. Biol."},{"key":"e_1_3_1_10_2","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1016\/S1097-2765(00)00047-2","volume":"6","author":"Demeter J.","year":"2000","unstructured":"Demeter J., Lee S. E., Haber J. E., Stearns T., Mol. Cell 6, 487 (2000).","journal-title":"Mol. Cell"},{"key":"e_1_3_1_11_2","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1016\/0076-6879(91)94011-Z","volume":"194","author":"Herskowitz I.","year":"1991","unstructured":"Herskowitz I., Jensen R. E., Methods Enzymol. 194, 132 (1991).","journal-title":"Methods Enzymol."},{"key":"e_1_3_1_12_2","unstructured":"All strains were isogenic with KSC006 and were derived using standard genetic techniques. Strains are MATa except for those indicated as MAT \u03b1. To express Rad53-HA cells were transformed with YCpT-RAD53-HA a TRP1 marker version of YCpRAD53-HA (16)."},{"key":"e_1_3_1_13_2","unstructured":"T. Kondo T. Wakayama T. Naiki K. Matsumoto K. Sugimoto unpublished data."},{"key":"e_1_3_1_14_2","first-page":"2342","volume":"8","author":"Connolly B.","year":"1988","unstructured":"Connolly B., White C. I., Haber J. E., Mol. Cell. Biol. 8, 2342 (1988).","journal-title":"Mol. Cell. Biol."},{"key":"e_1_3_1_15_2","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1016\/S1097-2765(01)00177-0","volume":"7","author":"Pellicioli A.","year":"2001","unstructured":"Pellicioli A., Lee S. E., Lucca C., Foiani M., Haber J. E., Mol. Cell 7, 293 (2001).","journal-title":"Mol. Cell"},{"key":"e_1_3_1_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1097-2765(00)80277-4"},{"key":"e_1_3_1_17_2","doi-asserted-by":"crossref","first-page":"1136","DOI":"10.1128\/MCB.19.2.1136","volume":"19","author":"Kondo T.","year":"1999","unstructured":"Kondo T., Matsumoto K., Sugimoto K., Mol. Cell. Biol. 19, 1136 (1999).","journal-title":"Mol. Cell. Biol."},{"key":"e_1_3_1_18_2","unstructured":"Yeast cells were cultured in medium containing 2% sucrose and then one-half of the culture was maintained in sucrose and the other half was incubated with 2% galactose to induce HO expression. Cells were converted to spheroplasts with the use of oxalyticase (50 \u03bcg\/ml) in 50 mM Hepes-KOH (pH 7.5); 100 mM NaCl; 0.4 M sorbitol; 50 mM NaF; 5 mM PNPP; 1 mM Na3VO4; 0.5 mM PMSF; aprotinin (13 \u03bcg\/ml); leupeptin (5 \u03bcg\/ml); pepstatin (5 \u03bcg\/ml); and 1 mM benzamidine. Spheroplasts were lysed by adding Triton X-100 (0.25%) and were subsequently cross-linked with 1% formaldehyde for 15 min at 4\u00b0C. Immunoprecipitation of cross-linked DNA was performed essentially as described (29) with the use of monoclonal antibody 12CA5 to HA. DNA was also purified from the whole cell extract (designated \u201cinput\u201d). PCR was done in 50-\u03bcl volumes containing 1\/25 of the anti-HA antibody immunoprecipitates or 1\/1200 of input respectively. The PCR products were separated in 2% agarose gels and were stained with ethidium bromide. The sequences of primers for the are as follows: HO1 set at MAT \u03b1 locus 5\u2032-CCAGATTTGTATTAGACGAGGGACGGAGTGA-3\u2032 and 5\u2032-AGAGGGTCACAGCACTAATACAGCTCGTAAT-3\u2032; HO2 set at MAT \u03b1 locus 5\u2032-GGTGTCCTCTGTAAGGTTTAGTACTTTTGT-3\u2032 and 5\u2032-CACAGATGAGTTTAAATCCAGCATACTAGACA-3\u2032; and SMC2 locus 5\u2032-AAGAGAAACTTTAGTCAAAACATGGG-3\u2032 and 5\u2032-CCATCACATTATACTAACTACGG-3\u2032."},{"key":"e_1_3_1_19_2","unstructured":"Cells were processed and incubated with mouse antibody to HA (16B12; Babco Richmond CA). Antibody binding was detected using Cy3-conjugated anti-mouse antibody (Jackson ImmunoResearch West Grove PA)."},{"key":"e_1_3_1_20_2","doi-asserted-by":"crossref","first-page":"4199","DOI":"10.1093\/emboj\/17.14.4199","volume":"17","author":"Paciotti V.","year":"1998","unstructured":"Paciotti V., Lucchini G., Plevani P., Longhese M. P., EMBO J. 17, 4199 (1998).","journal-title":"EMBO J."},{"key":"e_1_3_1_21_2","doi-asserted-by":"crossref","first-page":"5485","DOI":"10.1128\/MCB.18.9.5485","volume":"18","author":"Shimomura T.","year":"1998","unstructured":"Shimomura T., Ando S., Matsumoto K., Sugimoto K., Mol. Cell. Biol. 18, 5485 (1998).","journal-title":"Mol. Cell. Biol."},{"key":"e_1_3_1_22_2","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/S0960-9822(99)00263-8","volume":"10","author":"Green C. M.","year":"2000","unstructured":"Green C. M., Erdjument-Bromage H., Tempst P., Lowndes N. F., Curr. Biol. 10, 39 (2000).","journal-title":"Curr. 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Chem."},{"key":"e_1_3_1_29_2","doi-asserted-by":"crossref","first-page":"2989","DOI":"10.1101\/gad.851000","volume":"14","author":"Tibbetts R. S.","year":"2000","unstructured":"Tibbetts R. S., et al., Genes Dev. 14, 2989 (2000).","journal-title":"Genes Dev."},{"key":"e_1_3_1_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)80526-7"},{"key":"e_1_3_1_31_2","unstructured":"We thank M. P. Longhese G. Lucchini and T. Weinert for materials; we also thank J. Haber H. Araki M. Lamphier N. F. Lowndes and H. Masumoto for critical reading discussion and suggestion. Supported by grants from the Ministry of Education Science Sports and Culture of Japan."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.1063827","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,9]],"date-time":"2024-01-09T16:58:18Z","timestamp":1704819498000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.1063827"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2001,10,26]]},"references-count":30,"journal-issue":{"issue":"5543","published-print":{"date-parts":[[2001,10,26]]}},"alternative-id":["10.1126\/science.1063827"],"URL":"https:\/\/doi.org\/10.1126\/science.1063827","relation":{"has-review":[{"id-type":"doi","id":"10.3410\/f.1002056.19304","asserted-by":"object"},{"id-type":"doi","id":"10.3410\/f.1002056.20606","asserted-by":"object"},{"id-type":"doi","id":"10.3410\/f.1002056.26556","asserted-by":"object"},{"id-type":"doi","id":"10.3410\/f.1002056.17108","asserted-by":"object"}]},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2001,10,26]]}}}