{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T23:20:43Z","timestamp":1789600843966,"version":"build-2803163510"},"reference-count":40,"publisher":"American Chemical Society (ACS)","issue":"6","license":[{"start":{"date-parts":[[2020,9,22]],"date-time":"2020-09-22T00:00:00Z","timestamp":1600732800000},"content-version":"vor","delay-in-days":368,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_termsofuse.html"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["16H05346"],"award-info":[{"award-number":["16H05346"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/na","name":"Research Center for Zoonosis Control, Hokkaido University","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,6,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Multidrug-resistant (MDR) bacteria are widespread throughout the world and pose an increasingly serious threat to human and animal health. Besides implementing strict measures to prevent improper antibiotic use, it remains essential that novel antibiotics must be developed. These antibiotics need to exert their activity via mechanisms different from those employed by currently approved antibiotics. In this study, we used several 5-fluorouracil (5-FU) analogues as chemical probes and investigated the potential of these pyrimidine analogues as antibacterial agents. Several 5-FU derivatives exerted potent activity against strains of Gram-positive cocci (GPC) that are susceptible or resistant toward approved antibiotics, without showing cross-resistance. Furthermore, we have provided evidence that the pyrimidine analogues exerted anti-GPC activity via thymineless death by inhibition of thymidylate synthetase (ThyA) and\/or inhibition of RNA synthesis. Interestingly, whole genome resequencing of in vitro-selected, pyrimidine analogue-resistant Staphylococcus aureus mutants indicated that S.\u00a0aureus strains with pyrimidine-analogue resistance induced an amino acid (AA) substitution, deletion, and\/or insertion into thymineless-death related proteins except for ThyA, or enhanced the ThyA transcription level. Thus, S.\u00a0aureus may avoid altering the ThyA function by introducing an AA substitution, suggesting that the pyrimidine analogues, which directly bind to ThyA without phosphorylation, may be more effective and show a higher genetic barrier than the pyrimidines that depend on phosphorylation for activity. The findings of this study may assist in the future development of a novel class of antibiotics for combating MDR GPC, including methicillin-resistant S.\u00a0aureus and vancomycin-resistant Enterococci.<\/jats:p>","DOI":"10.1021\/acsinfecdis.9b00305","type":"journal-article","created":{"date-parts":[[2019,9,20]],"date-time":"2019-09-20T21:07:21Z","timestamp":1569013641000},"page":"1490-1500","source":"Crossref","is-referenced-by-count":30,"title":["Pyrimidine Analogues as a New Class of Gram-Positive\nAntibiotics, Mainly Targeting Thymineless-Death Related Proteins"],"prefix":"10.1021","volume":"6","author":[{"given":"Chihiro","family":"Oe","sequence":"first","affiliation":[{"name":"Tohoku University Graduate School of Medicine , , 2-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8575"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5892-0040","authenticated-orcid":true,"given":"Hironori","family":"Hayashi","sequence":"additional","affiliation":[{"name":"Tohoku University Hospital , , 1-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8574"]},{"name":"Tohoku University Graduate School of Medicine , , 2-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8575"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazushige","family":"Hirata","sequence":"additional","affiliation":[{"name":"Tohoku University Hospital , , 1-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8574"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kumi","family":"Kawaji","sequence":"additional","affiliation":[{"name":"Tohoku University , , 2-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8575"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fusako","family":"Hashima","sequence":"additional","affiliation":[{"name":"Tohoku University Hospital , , 1-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8574"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mina","family":"Sasano","sequence":"additional","affiliation":[{"name":"Tohoku University , , 2-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8575"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maaya","family":"Furuichi","sequence":"additional","affiliation":[{"name":"Tohoku University , , 2-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8575"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emiko","family":"Usui","sequence":"additional","affiliation":[{"name":"Tohoku University , , 2-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8575"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Makoto","family":"Katsumi","sequence":"additional","affiliation":[{"name":"Tohoku University Hospital , , 1-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8574"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasuhiko","family":"Suzuki","sequence":"additional","affiliation":[{"name":"Hokkaido University Research Centre for Zoonosis Control , , North 20, West 10 Kita-ku , , ,","place":["Sapporo, Hokkaido, Japan, 001-0020"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chie","family":"Nakajima","sequence":"additional","affiliation":[{"name":"Hokkaido University Research Centre for Zoonosis Control , , North 20, West 10 Kita-ku , , ,","place":["Sapporo, Hokkaido, Japan, 001-0020"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mitsuo","family":"Kaku","sequence":"additional","affiliation":[{"name":"Tohoku University Graduate School of Medicine , , 2-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8575"]},{"name":"Tohoku University Hospital , , 1-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8574"]},{"name":"Tohoku University Graduate School of Medicine , , 2-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8575"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eiichi N.","family":"Kodama","sequence":"additional","affiliation":[{"name":"Tohoku University , , 2-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8575"]},{"name":"Tohoku University , , 2-1, Seiryo-machi, Aoba-ku , , ,","place":["Sendai, Miyagi, Japan, 980-8575"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2019,9,20]]},"reference":[{"key":"2026090120515703600_cit1","doi-asserted-by":"publisher","first-page":"529","DOI":"10.1038\/nrd4572","article-title":"ESKAPEing the labyrinth of antibacterial\ndiscovery","volume":"14","author":"Tommasi","year":"2015","journal-title":"Nat. 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Bacteriol."},{"issue":"4","key":"2026090120515703600_cit20","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01447-14","article-title":"Inactivation of thyA in Staphylococcus aureus attenuates\nvirulence and has a strong impact on metabolism and virulence gene\nexpression","volume":"5","author":"Kriegeskorte","year":"2014","journal-title":"mBio"},{"key":"2026090120515703600_cit21","volume-title":"Performance Standards for Antimicrobial\nSusceptibility Testing (M100S26)","author":"Clinical and Laboratory Standards Institute (CLSI).","year":"2016"},{"issue":"1","key":"2026090120515703600_cit22","doi-asserted-by":"publisher","first-page":"134","DOI":"10.1111\/1574-6968.12491","article-title":"Validation of reference\ngenes for normalization of qPCR mRNA expression levels in Staphylococcus\naureus exposed to osmotic and lactic acid stress conditions encountered\nduring food production and preservation","volume":"356","author":"Sihto","year":"2014","journal-title":"FEMS\nMicrobiol. Lett."},{"issue":"7","key":"2026090120515703600_cit23","doi-asserted-by":"crossref","first-page":"1258","DOI":"10.1128\/jcm.25.7.1258-1261.1987","article-title":"Prevalence of thymidine-dependent Staphylococcus aureus\nin patients with cystic fibrosis","volume":"25","author":"Gilligan","year":"1987","journal-title":"J. Clin Microbiol"},{"key":"2026090120515703600_cit24","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1016\/j.mimet.2015.01.007","article-title":"Addition of thymidine to culture media for accurate examination of\nthymidine-dependent small-colony variants of methicillin-resistant\nStaphylococcus aureus: a pilot study","volume":"110","author":"Horiuchi","year":"2015","journal-title":"J. Microbiol.\nMethods"},{"issue":"1","key":"2026090120515703600_cit25","doi-asserted-by":"publisher","first-page":"62","DOI":"10.1111\/j.1751-7915.2008.00060.x","article-title":"Uracil influences\nquorum sensing and biofilm formation in Pseudomonas aeruginosa and\nfluorouracil is an antagonist","volume":"2","author":"Ueda","year":"2009","journal-title":"Microb. Biotechnol."},{"issue":"1","key":"2026090120515703600_cit26","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1053\/ctrv.2002.0253","article-title":"Thymidylate synthase expression as\na predictor of clinical response to fluoropyrimidine-based chemotherapy\nin advanced colorectal cancer","volume":"28","author":"Aschele","year":"2002","journal-title":"Cancer Treat.\nRev."},{"issue":"4","key":"2026090120515703600_cit27","first-page":"1149","article-title":"Chronic exposure of colorectal cancer cells in culture\nto fluoropyrimidine analogs induces thymidylate synthase and suppresses\np53. A molecular explanation for the mechanism of 5-FU resistance","volume":"30","author":"Subbarayan","year":"2010","journal-title":"Anticancer Res."},{"issue":"12","key":"2026090120515703600_cit28","doi-asserted-by":"publisher","first-page":"7265","DOI":"10.1128\/AAC.00742-15","article-title":"Thymidine-Dependent Staphylococcus aureus Small-Colony\nVariants Are Induced by Trimethoprim-Sulfamethoxazole (SXT) and Have\nIncreased Fitness during SXT Challenge","volume":"59","author":"Kriegeskorte","year":"2015","journal-title":"Antimicrob.\nAgents Chemother."},{"issue":"9","key":"2026090120515703600_cit29","doi-asserted-by":"publisher","first-page":"1893","DOI":"10.1080\/00021369.1976.10862322","article-title":"Distribution\nof enzymes related to cytidine degradation in bacteria","volume":"40","author":"Sakai","year":"1976","journal-title":"Agric. Biol. Chem."},{"issue":"8","key":"2026090120515703600_cit30","doi-asserted-by":"publisher","first-page":"559","DOI":"10.1038\/nrd.2018.46","article-title":"The expanding role of prodrugs in contemporary drug\ndesign and development","volume":"17","author":"Rautio","year":"2018","journal-title":"Nat. Rev. Drug Discovery"},{"key":"2026090120515703600_cit31","doi-asserted-by":"publisher","first-page":"810","DOI":"10.1016\/j.ejmech.2016.10.061","article-title":"Prodrug approach: An overview of\nrecent cases","volume":"127","author":"Abet","year":"2017","journal-title":"Eur. J. Med. Chem."},{"issue":"11","key":"2026090120515703600_cit32","doi-asserted-by":"publisher","first-page":"4733","DOI":"10.1128\/AAC.00605-10","article-title":"Mutational analysis\nof flucytosine resistance in Candida glabrata","volume":"54","author":"Edlind","year":"2010","journal-title":"Antimicrob. Agents Chemother."},{"key":"2026090120515703600_cit33","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1016\/j.antiviral.2015.11.009","article-title":"Tenofovir\nalafenamide: a novel prodrug of tenofovir for the treatment of human\nimmunodeficiency virus","volume":"125","author":"Ray","year":"2016","journal-title":"Antiviral Res."},{"key":"2026090120515703600_cit34","doi-asserted-by":"publisher","first-page":"218","DOI":"10.1016\/j.molcatb.2016.08.015","article-title":"New developments in nucleoside analogues\nbiosynthesis: A review","volume":"133","author":"Lapponi","year":"2016","journal-title":"J. Mol. Catal. B: Enzym."},{"issue":"8","key":"2026090120515703600_cit35","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00620-18","article-title":"Nucleotide prodrug\ncontaining a nonproteinogenic amino acid to improve oral delivery\nof a hepatitis C virus treatment","volume":"62","author":"Feng","year":"2018","journal-title":"Antimicrob.\nAgents Chemother."},{"issue":"28","key":"2026090120515703600_cit36","doi-asserted-by":"publisher","first-page":"7720","DOI":"10.1039\/C5OB01037C","article-title":"Design, synthesis\nand biological evaluation of 5\u2032-C-piperidinyl-5\u2032-O-aminoribosyluridines\nas potential antibacterial agents","volume":"13","author":"Nakaya","year":"2015","journal-title":"Org. Biomol.\nChem."},{"issue":"12","key":"2026090120515703600_cit37","doi-asserted-by":"publisher","first-page":"2651","DOI":"10.1021\/bi00307a017","article-title":"Alterations in the maturation and structure of ribosomal\nprecursor RNA in Novikoff hepatoma cells induced by 5-fluorocytidine","volume":"23","author":"Grosso","year":"1984","journal-title":"Biochemistry"},{"issue":"11","key":"2026090120515703600_cit38","first-page":"3014","article-title":"The inhibition of ribosomal RNA synthesis\nand maturation in Novikoff hepatoma cells by 5-fluorouridine","volume":"35","author":"Wilkinson","year":"1975","journal-title":"Cancer Res."},{"key":"2026090120515703600_cit39","doi-asserted-by":"publisher","first-page":"247","DOI":"10.1146\/annurev-micro-092412-155749","article-title":"Thymineless Death Lives On: New Insights\ninto a Classic Phenomenon","volume":"69","author":"Khodursky","year":"2015","journal-title":"Annu. Rev. Microbiol."},{"issue":"9","key":"2026090120515703600_cit40","doi-asserted-by":"publisher","first-page":"1286","DOI":"10.2174\/092986711795029555","article-title":"Pyrimidine salvage\npathway in Mycobacterium tuberculosis","volume":"18","author":"Villela","year":"2011","journal-title":"Curr.\nMed. Chem."}],"container-title":["ACS Infectious Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acsinfecdis.9b00305","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article-pdf\/6\/6\/1490\/67570616\/acsinfecdis.9b00305.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article-pdf\/6\/6\/1490\/67570616\/acsinfecdis.9b00305.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T00:52:02Z","timestamp":1788310322000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article\/6\/6\/1490\/607864\/Pyrimidine-Analogues-as-a-New-Class-of-Gram"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,9,20]]},"references-count":40,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2019,9,20]]},"published-print":{"date-parts":[[2020,6,12]]}},"URL":"https:\/\/doi.org\/10.1021\/acsinfecdis.9b00305","relation":{},"ISSN":["2373-8227"],"issn-type":[{"value":"2373-8227","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,9,20]]}}}