{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T01:21:30Z","timestamp":1791940890348,"version":"4.3.4"},"reference-count":36,"publisher":"National Academy of Sciences","issue":"15","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2000,7,18]]},"abstract":"<jats:p>\n                    Three cytosolic and one plasma membrane-bound 5\u2032-nucleotidases have been cloned and characterized. Their various substrate specificities suggest widely different functions in nucleotide metabolism. We now describe a 5\u2032-nucleotidase in mitochondria. The enzyme, named dNT-2, dephosphorylates specifically the 5\u2032- and 2\u2032(3\u2032)-phosphates of uracil and thymine deoxyribonucleotides. The cDNA of human dNT-2 codes for a 25.9-kDa polypeptide with a typical mitochondrial leader peptide, providing the structural basis for two-step processing during import into the mitochondrial matrix. The deduced amino acid sequence is 52% identical to that of a recently described cytosolic deoxyribonucleotidase (dNT-1). The two enzymes share many catalytic properties, but dNT-2 shows a narrower substrate specificity. Mitochondrial localization of dNT-2 was demonstrated by the mitochondrial fluorescence of 293 cells expressing a dNT-2-green fluorescent protein (GFP) fusion protein. 293 cells expressing fusion proteins without leader peptide or with dNT-1 showed a cytosolic fluorescence. During\n                    <jats:italic>in vitro<\/jats:italic>\n                    import into mitochondria, the preprotein lost the leader peptide. We suggest that dNT-2 protects mitochondrial DNA replication from overproduction of dTTP, in particular in resting cells. Mitochondrial toxicity of dTTP can be inferred from a severe inborn error of metabolism in which the loss of thymidine phosphorylase led to dTTP accumulation and aberrant mitochondrial DNA replication. We localized the gene for dNT-2 on chromosome 17p11.2 in the Smith\u2013Magenis syndrome-critical region, raising the possibility that dNT-2 is involved in the etiology of this genetic disease.\n                  <\/jats:p>","DOI":"10.1073\/pnas.97.15.8239","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T10:41:48Z","timestamp":1027680108000},"page":"8239-8244","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":79,"title":["A deoxyribonucleotidase in mitochondria: Involvement in regulation of dNTP pools and possible link to genetic disease"],"prefix":"10.1073","volume":"97","author":[{"given":"Chiara","family":"Rampazzo","sequence":"first","affiliation":[{"name":"Departments of Biology and Biomedical Sciences, University of Padua, I-35131 Padua, Italy; and Department of Biochemistry, Karolinska Institutet, S-17177 Stockholm, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa","family":"Gallinaro","sequence":"additional","affiliation":[{"name":"Departments of Biology and Biomedical Sciences, University of Padua, I-35131 Padua, Italy; and Department of Biochemistry, Karolinska Institutet, S-17177 Stockholm, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Milanesi","sequence":"additional","affiliation":[{"name":"Departments of Biology and Biomedical Sciences, University of Padua, I-35131 Padua, Italy; and Department of Biochemistry, Karolinska Institutet, S-17177 Stockholm, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisabetta","family":"Frigimelica","sequence":"additional","affiliation":[{"name":"Departments of Biology and Biomedical Sciences, University of Padua, I-35131 Padua, Italy; and Department of Biochemistry, Karolinska Institutet, S-17177 Stockholm, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Reichard","sequence":"additional","affiliation":[{"name":"Departments of Biology and Biomedical Sciences, University of Padua, I-35131 Padua, Italy; and Department of Biochemistry, Karolinska Institutet, S-17177 Stockholm, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vera","family":"Bianchi","sequence":"additional","affiliation":[{"name":"Departments of Biology and Biomedical Sciences, University of Padua, I-35131 Padua, Italy; and Department of Biochemistry, Karolinska Institutet, S-17177 Stockholm, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2000,7,18]]},"reference":[{"key":"e_1_3_4_1_2","first-page":"680","volume-title":"DNA Replication","author":"Kornberg A","year":"1992","unstructured":"A Kornberg, T A Baker DNA Replication (Freeman, New York,), pp. 680\u2013687 (1992)."},{"key":"e_1_3_4_2_2","doi-asserted-by":"crossref","first-page":"2722","DOI":"10.1016\/S0021-9258(19)44066-0","volume":"248","author":"Berk A J","year":"1973","unstructured":"A J Berk, D A Clayton J Biol Chem 248, 2722\u20132729 (1973).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_3_2","doi-asserted-by":"crossref","first-page":"9300","DOI":"10.1016\/S0021-9258(18)34068-7","volume":"257","author":"Bestwick R K","year":"1982","unstructured":"R K Bestwick, G L Moeffett, C K Mathews J Biol Chem 257, 9300\u20139304 (1982).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_4_2","doi-asserted-by":"crossref","first-page":"4620","DOI":"10.1074\/jbc.274.8.4620","volume":"274","author":"Bridges E G","year":"1999","unstructured":"E G Bridges, Z Jiang, Y-C Cheng J Biol Chem 274, 4620\u20134625 (1999).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_5_2","doi-asserted-by":"crossref","first-page":"8454","DOI":"10.1074\/jbc.272.13.8454","volume":"272","author":"Johansson M","year":"1997","unstructured":"M Johansson, A Karlsson J Biol Chem 272, 8454\u20138458 (1997).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_6_2","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1016\/S0014-5793(98)01711-6","volume":"443","author":"Wang L","year":"1999","unstructured":"L Wang, A Herrstr\u00f6m Sj\u00f6berg, T Bergman, U Hellman, S Eriksson FEBS Lett 443, 170\u2013174 (1999).","journal-title":"FEBS Lett"},{"key":"e_1_3_4_7_2","doi-asserted-by":"crossref","first-page":"7258","DOI":"10.1073\/pnas.93.14.7258","volume":"93","author":"Johansson M","year":"1996","unstructured":"M Johansson, A Karlsson Proc Natl Acad Sci USA 93, 7258\u20137262 (1996).","journal-title":"Proc Natl Acad Sci USA"},{"key":"e_1_3_4_8_2","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/0014-5793(96)00623-0","volume":"390","author":"Wang L","year":"1996","unstructured":"L Wang, U Hellman, S Eriksson FEBS Lett 390, 39\u201343 (1996).","journal-title":"FEBS Lett"},{"key":"e_1_3_4_9_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0165-1110(94)90006-X","volume":"318","author":"Kunz B A","year":"1994","unstructured":"B A Kunz, S E Kohalmi, T A Kunkel, C K Mathews, E M McIntosh, J A Reidy Mutation Res 318, 1\u201364 (1994).","journal-title":"Mutation Res"},{"key":"e_1_3_4_10_2","doi-asserted-by":"crossref","first-page":"1067","DOI":"10.1016\/S0140-6736(72)92345-8","author":"Giblett E R","year":"1972","unstructured":"E R Giblett, J E Anderson, F Cohen, B Pollara, H J Meuwissen Lancet ii, 1067\u20131069 (1972).","journal-title":"Lancet"},{"key":"e_1_3_4_11_2","doi-asserted-by":"crossref","first-page":"1010","DOI":"10.1016\/S0140-6736(75)91950-9","volume":"1","author":"Giblett E R","year":"1975","unstructured":"E R Giblett, A J Ammann, R Sandman, D W Wara, L K Diamond Lancet 1, 1010\u20131013 (1975).","journal-title":"Lancet"},{"key":"e_1_3_4_12_2","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1073\/pnas.75.1.472","volume":"75","author":"Cohen A","year":"1978","unstructured":"A Cohen, R Hirschhorn, S D Horowitz, A Rubistein, S H Polmar, R Hong, D W Martin Proc Natl Acad Sci USA 75, 472\u2013476 (1978).","journal-title":"Proc Natl Acad Sci USA"},{"key":"e_1_3_4_13_2","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.1172\/JCI109058","volume":"61","author":"Cohen A","year":"1978","unstructured":"A Cohen, L J Gudas, A J Ammann, G E Staal, D W Martin J Clin Invest 61, 1405\u20131409 (1978).","journal-title":"J Clin Invest"},{"key":"e_1_3_4_14_2","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1126\/science.283.5402.689","volume":"283","author":"Nishino I","year":"1999","unstructured":"I Nishino, A Spinazzola, M Hirano Science 283, 689\u2013692 (1999).","journal-title":"Science"},{"key":"e_1_3_4_15_2","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1146\/annurev.biochem.67.1.71","volume":"67","author":"Jordan A","year":"1998","unstructured":"A Jordan, P Reichard Annu Rev Biochem 67, 71\u201398 (1998).","journal-title":"Annu Rev Biochem"},{"key":"e_1_3_4_16_2","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1146\/annurev.bi.57.070188.002025","volume":"57","author":"Reichard P","year":"1988","unstructured":"P Reichard Annu Rev Biochem 57, 349\u2013374 (1988).","journal-title":"Annu Rev Biochem"},{"key":"e_1_3_4_17_2","doi-asserted-by":"crossref","first-page":"348","DOI":"10.4049\/jimmunol.126.1.348","volume":"126","author":"Carson D","year":"1981","unstructured":"D Carson, J Kaye, D B Wasson J Immunol 126, 348\u2013352 (1981).","journal-title":"J Immunol"},{"key":"e_1_3_4_18_2","first-page":"3681","volume":"48","author":"H\u00f6glund L","year":"1988","unstructured":"L H\u00f6glund, E Pontis, P Reichard Cancer Res 48, 3681\u20133687 (1988).","journal-title":"Cancer Res"},{"key":"e_1_3_4_19_2","doi-asserted-by":"crossref","first-page":"16677","DOI":"10.1016\/S0021-9258(19)89443-7","volume":"269","author":"Bianchi V","year":"1994","unstructured":"V Bianchi, P Ferraro, S Borella, P Bonvini, P Reichard J Biol Chem 269, 16677\u201316683 (1994).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_20_2","doi-asserted-by":"crossref","first-page":"5409","DOI":"10.1074\/jbc.275.8.5409","volume":"275","author":"Rampazzo C","year":"2000","unstructured":"C Rampazzo, M Johansson, L Gallinaro, P Ferraro, U Hellman, A Karlsson, P Reichard, V Bianchi J Biol Chem 275, 5409\u20135415 (2000).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_21_2","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1002\/ajmg.1320240303","volume":"24","author":"Smith A C M","year":"1986","unstructured":"A C M Smith, L McGavran, J Robinson, G Waldstein, J Macfarlane, J Zonana, J Reiss, M Lahr, L Allen, E Magenis Am J Med Genet 24, 393\u2013414 (1986).","journal-title":"Am J Med Genet"},{"key":"e_1_3_4_22_2","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1006\/excr.1999.4681","volume":"253","author":"Gazziola C","year":"1999","unstructured":"C Gazziola, M Moras, P Ferraro, L Gallinaro, R Verin, C Rampazzo, P Reichard, V Bianchi Exp Cell Res 253, 474\u2013482 (1999).","journal-title":"Exp Cell Res"},{"key":"e_1_3_4_23_2","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1083\/jcb.32.2.415","volume":"32","author":"Sottocasa G L","year":"1967","unstructured":"G L Sottocasa, B Kuylestierna, L Ernster, A Bergstrand J Cell Biol 32, 415\u2013438 (1967).","journal-title":"J Cell Biol"},{"key":"e_1_3_4_24_2","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1016\/S0091-679X(08)61689-1","volume":"34","author":"S\u00f6llner T","year":"1991","unstructured":"T S\u00f6llner, J Rassow, N Pfanner Methods Cell Biol 34, 345\u2013358 (1991).","journal-title":"Methods Cell Biol"},{"key":"e_1_3_4_25_2","doi-asserted-by":"crossref","first-page":"863","DOI":"10.1146\/annurev.biochem.66.1.863","volume":"66","author":"Neupert W","year":"1997","unstructured":"W Neupert Annu Rev Biochem 66, 863\u2013917 (1997).","journal-title":"Annu Rev Biochem"},{"key":"e_1_3_4_26_2","doi-asserted-by":"crossref","first-page":"27366","DOI":"10.1074\/jbc.270.45.27366","volume":"270","author":"Branda S S","year":"1995","unstructured":"S S Branda, G Isaya J Biol Chem 270, 27366\u201327373 (1995).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_27_2","doi-asserted-by":"crossref","first-page":"2936","DOI":"10.1074\/jbc.272.5.2936","volume":"272","author":"Spyrou G","year":"1997","unstructured":"G Spyrou, E Enmark, A Miranda-Vizuete, J-\u00c5 Gustafsson J Biol Chem 272, 2936\u20132941 (1997).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_28_2","doi-asserted-by":"crossref","first-page":"4722","DOI":"10.1074\/jbc.274.8.4722","volume":"274","author":"Lee S-R","year":"1999","unstructured":"S-R Lee, J R Kim, K S Kwon, H W Yoon, R L Levine, A Ginsburg, S G Rhee J Biol Chem 274, 4722\u20134734 (1999).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_29_2","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1006\/geno.1998.5748","volume":"57","author":"Potocki L","year":"1999","unstructured":"L Potocki, K-S Chen, J R Lupski Genomics 57, 180\u2013182 (1999).","journal-title":"Genomics"},{"key":"e_1_3_4_30_2","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1002\/(SICI)1098-2779(1996)2:3<122::AID-MRDD2>3.0.CO;2-U","volume":"2","author":"Chen K-S","year":"1996","unstructured":"K-S Chen, L Potocki, J R Lupski Ment Retard Dev Disabil Res Rev 2, 122\u2013129 (1996).","journal-title":"Ment Retard Dev Disabil Res Rev"},{"key":"e_1_3_4_31_2","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1146\/annurev.biochem.67.1.509","volume":"67","author":"Tsien R","year":"1998","unstructured":"R Tsien Annu Rev Biochem 67, 509\u2013544 (1998).","journal-title":"Annu Rev Biochem"},{"key":"e_1_3_4_32_2","doi-asserted-by":"crossref","first-page":"3346","DOI":"10.1073\/pnas.93.8.3346","volume":"93","author":"No D","year":"1996","unstructured":"D No, T-P Yao, R M Evans Proc Natl Acad Sci USA 93, 3346\u20133351 (1996).","journal-title":"Proc Natl Acad Sci USA"},{"key":"e_1_3_4_33_2","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1042\/bj2850345","volume":"285","author":"Zimmermann H","year":"1992","unstructured":"H Zimmermann Biochem J 285, 345\u2013365 (1992).","journal-title":"Biochem J"},{"key":"e_1_3_4_34_2","first-page":"13","volume":"105","author":"Itoh R","year":"1992","unstructured":"R Itoh Comp Biochem Physiol 105B, 13\u201319 (1992).","journal-title":"Comp Biochem Physiol"},{"key":"e_1_3_4_35_2","doi-asserted-by":"crossref","first-page":"9114","DOI":"10.1016\/S0021-9258(17)39337-7","volume":"260","author":"Engstr\u00f6m Y","year":"1985","unstructured":"Y Engstr\u00f6m, S Eriksson, I Jildevik, S Skog, L Thelander, B Tribukait J Biol Chem 260, 9114\u20139116 (1985).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_36_2","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1038\/ng1097-154","volume":"17","author":"Chen K-S","year":"1997","unstructured":"K-S Chen, P Manian, T Koeuth, L Potocki, Q Zhao, A C Chinault, C C Lee, J R Lupski Nat Genet 17, 154\u2013163 (1997).","journal-title":"Nat Genet"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.97.15.8239","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,4,22]],"date-time":"2023-04-22T08:27:26Z","timestamp":1682152046000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.97.15.8239"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2000,7,18]]},"references-count":36,"journal-issue":{"issue":"15","published-print":{"date-parts":[[2000,7,18]]}},"alternative-id":["10.1073\/pnas.97.15.8239"],"URL":"https:\/\/doi.org\/10.1073\/pnas.97.15.8239","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2000,7,18]]},"assertion":[{"value":"2000-05-18","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2000-07-18","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}