{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T08:06:00Z","timestamp":1790409960540,"version":"4.1.0"},"reference-count":36,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2009,4,1]],"date-time":"2009-04-01T00:00:00Z","timestamp":1238544000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Bacteriol"],"published-print":{"date-parts":[[2009,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Thiamine pyrophosphate is an essential cofactor involved in central metabolism and amino acid biosynthesis and is derived from thiamine (vitamin B<jats:sub>1<\/jats:sub>). The extent to which this metabolite is available to bacterial pathogens replicating within host cells is still little understood. Growth studies using modified minimal Welshimer\u2032s broth (mMWB) supplemented with thiamine or the thiamine precursor hydroxymethylpyrimidine (HMP) showed that<jats:italic>Listeria monocytogenes<\/jats:italic>, in agreement with bioinformatic prediction, is able to synthesize thiamine only in the presence of HMP. This appears to be due to a lack of ThiC, which is involved in HMP synthesis. The knockout of<jats:italic>thiD<\/jats:italic>(lmo0317), which probably catalyzes the phosphorylation of HMP, inhibited growth in mMWB supplemented with HMP and reduced the replication rate of<jats:italic>L. monocytogenes<\/jats:italic>in epithelial cells. Mutation of a predicted thiamine transporter gene, lmo1429, led to reduced proliferation of<jats:italic>L. monocytogenes<\/jats:italic>in mMWB containing thiamine or thiamine phosphates and also within epithelial cells but had no influence on the expression of the virulence factors Hly and ActA. The toxic thiamine analogue pyrithiamine inhibited growth of wild-type strain EGD but not of the transporter mutant EGD\u0394<jats:italic>thiT<\/jats:italic>. We also demonstrated that ThiT binds thiamine, a finding compatible with ThiT acting as the substrate-binding component of a multimeric thiamine transporter complex. These data provide experimental evidence that Lmo1429 homologs including<jats:italic>Bacillus<\/jats:italic>YuaJ are necessary for thiamine transport in gram-positive bacteria and are therefore proposed to be annotated \u201cThiT.\u201d Taken together, these data indicate that concurrent thiamine uptake and biosynthesis of thiamine precursors is a strategy of<jats:italic>L. monocytogenes<\/jats:italic>and possibly other facultative intracellular pathogens to enable proliferation within the cytoplasm.<\/jats:p>","DOI":"10.1128\/jb.01636-08","type":"journal-article","created":{"date-parts":[[2009,1,31]],"date-time":"2009-01-31T01:45:42Z","timestamp":1233366342000},"page":"2218-2227","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":61,"title":["Both Thiamine Uptake and Biosynthesis of Thiamine Precursors Are Required for Intracellular Replication of<i>Listeria monocytogenes<\/i>"],"prefix":"10.1128","volume":"191","author":[{"given":"Kristina","family":"Schauer","sequence":"first","affiliation":[{"name":"Zentralinstitut fu\u0308r Erna\u0308hrungs- und Lebensmittelforschung (ZIEL), Abteilung Mikrobiologie"},{"name":"Lehrstuhl fu\u0308r Mikrobielle O\u0308kologie, Technische Universita\u0308t Mu\u0308nchen, Weihenstephaner Berg 3, and Am Forum 5, 85354 Freising, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ju\u0308rgen","family":"Stolz","sequence":"additional","affiliation":[{"name":"Abteilung Biochemie"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Siegfried","family":"Scherer","sequence":"additional","affiliation":[{"name":"Zentralinstitut fu\u0308r Erna\u0308hrungs- und Lebensmittelforschung (ZIEL), Abteilung Mikrobiologie"},{"name":"Lehrstuhl fu\u0308r Mikrobielle O\u0308kologie, Technische Universita\u0308t Mu\u0308nchen, Weihenstephaner Berg 3, and Am Forum 5, 85354 Freising, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thilo M.","family":"Fuchs","sequence":"additional","affiliation":[{"name":"Zentralinstitut fu\u0308r Erna\u0308hrungs- und Lebensmittelforschung (ZIEL), Abteilung Mikrobiologie"},{"name":"Lehrstuhl fu\u0308r Mikrobielle O\u0308kologie, Technische Universita\u0308t Mu\u0308nchen, Weihenstephaner Berg 3, and Am Forum 5, 85354 Freising, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"crossref","first-page":"6069","DOI":"10.1093\/nar\/18.20.6069","volume":"18","year":"1990","unstructured":"Aslanidis, C., and P. J. de Jong. 1990. Ligation-independent cloning of PCR products (LIC-PCR). Nucleic Acids Res. 18 : 6069-6074.","journal-title":"Nucleic Acids Res."},{"key":"e_1_3_2_3_2","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1038\/nature04616","volume":"440","year":"2006","unstructured":"Becker, D., M. Selbach, C. Rollenhagen, M. Ballmaier, T. F. Meyer, M. Mann, and D. Bumann. 2006. Robust Salmonella metabolism limits possibilities for new antimicrobials. Nature 440 : 303-307.","journal-title":"Nature"},{"key":"e_1_3_2_4_2","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1007\/s002030050713","volume":"171","year":"1999","unstructured":"Begley, T. P., D. M. Downs, S. E. Ealick, F. W. McLafferty, A. P. Van Loon, S. Taylor, N. Campobasso, H. J. Chiu, C. Kinsland, J. J. Reddick, and J. Xi. 1999. Thiamin biosynthesis in prokaryotes. Arch. Microbiol. 171 : 293-300.","journal-title":"Arch. Microbiol."},{"key":"e_1_3_2_5_2","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1046\/j.1365-2958.2002.02767.x","volume":"43","year":"2002","unstructured":"Bergmann, B., D. Raffelsbauer, M. Kuhn, M. Goetz, S. Hom, and W. Goebel. 2002. InlA- but not InlB-mediated internalization of Listeria monocytogenes by non-phagocytic mammalian cells needs the support of other internalins. Mol. Microbiol. 43 : 557-570.","journal-title":"Mol. Microbiol."},{"key":"e_1_3_2_6_2","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1016\/0197-0186(94)00123-C","volume":"26","year":"1995","unstructured":"Bettendorff, L. 1995. Thiamine homeostasis in neuroblastoma cells. Neurochem. Int. 26 : 295-302.","journal-title":"Neurochem. Int."},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.68.12.7069-7077.2000"},{"key":"e_1_3_2_8_2","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1073\/pnas.012363899","volume":"99","year":"2002","unstructured":"Chico-Calero, I., M. Suarez, B. Gonzalez-Zorn, M. Scortti, J. Slaghuis, W. Goebel, and J. A. Vazquez-Boland. 2002. Hpt, a bacterial homolog of the microsomal glucose-6-phosphate translocase, mediates rapid intracellular proliferation in Listeria. Proc. Natl. Acad. Sci. USA 99 : 431-436.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01070-08"},{"key":"e_1_3_2_10_2","doi-asserted-by":"crossref","first-page":"849","DOI":"10.1126\/science.1063447","volume":"294","year":"2001","unstructured":"Glaser, P., L. Frangeul, C. Buchrieser, C. Rusniok, A. Amend, F. Baquero, P. Berche, H. Bloecker, P. Brandt, T. Chakraborty, A. Charbit, F. Chetouani, E. Couve, A. de Daruvar, P. Dehoux, E. Domann, G. Dominguez-Bernal, E. Duchaud, L. Durant, O. Dussurget, K. D. Entian, H. Fsihi, F. Garcia-del Portillo, P. Garrido, L. Gautier, W. Goebel, N. Gomez-Lopez, T. Hain, J. Hauf, D. Jackson, L. M. Jones, U. Kaerst, J. Kreft, M. Kuhn, F. Kunst, G. Kurapkat, E. Madueno, A. Maitournam, J. M. Vicente, E. Ng, H. Nedjari, G. Nordsiek, S. Novella, B. de Pablos, J. C. Perez-Diaz, R. Purcell, B. Remmel, M. Rose, T. Schlueter, N. Simoes, A. Tierrez, J. A. Vazquez-Boland, H. Voss, J. Wehland, and P. Cossart. 2001. Comparative genomics of Listeria species. Science 294 : 849-852.","journal-title":"Science"},{"key":"e_1_3_2_11_2","doi-asserted-by":"crossref","first-page":"12221","DOI":"10.1073\/pnas.211106398","volume":"98","year":"2001","unstructured":"Goetz, M., A. Bubert, G. Wang, I. Chico-Calero, J. A. Vazquez-Boland, M. Beck, J. Slaghuis, A. A. Szalay, and W. Goebel. 2001. Microinjection and growth of bacteria in the cytosol of mammalian host cells. Proc. Natl. Acad. Sci. USA 98 : 12221-12226.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(83)80284-8"},{"key":"e_1_3_2_13_2","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1111\/j.1574-6976.1999.tb00392.x","volume":"23","year":"1999","unstructured":"Handfield, M., and R. C. Levesque. 1999. Strategies for isolation of in vivo expressed genes from bacteria. FEMS Microbiol. Rev. 23 : 69-91.","journal-title":"FEMS Microbiol. Rev."},{"key":"e_1_3_2_14_2","doi-asserted-by":"crossref","first-page":"2909","DOI":"10.1073\/pnas.0609905104","volume":"104","year":"2007","unstructured":"Hebbeln, P., D. A. Rodionov, A. Alfandega, and T. Eitinger. 2007. Biotin uptake in prokaryotes by solute transporters with an optional ATP-binding cassette-containing module. Proc. Natl. Acad. Sci. USA 104 : 2909-2914.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_15_2","doi-asserted-by":"crossref","first-page":"492","DOI":"10.1038\/nchembio.2007.13","volume":"3","year":"2007","unstructured":"Jenkins, A. H., G. Schyns, S. Potot, G. Sun, and T. P. Begley. 2007. A new thiamin salvage pathway. Nat. Chem. Biol. 3 : 492-497.","journal-title":"Nat. Chem. Biol."},{"key":"e_1_3_2_16_2","doi-asserted-by":"crossref","first-page":"1188","DOI":"10.1016\/j.micinf.2007.05.006","volume":"9","year":"2007","unstructured":"Joseph, B., and W. Goebel. 2007. Life of Listeria monocytogenes in the host cells' cytosol. Microbes Infect. 9 : 1188-1195.","journal-title":"Microbes Infect."},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.188.2.556-568.2006"},{"key":"e_1_3_2_18_2","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1111\/j.1574-6968.2001.tb10930.x","volume":"205","year":"2001","unstructured":"Joseph, P., J. R. Fantino, M. L. Herbaud, and F. Denizot. 2001. Rapid orientated cloning in a shuttle vector allowing modulated gene expression in Bacillus subtilis. FEMS Microbiol. Lett. 205 : 91-97.","journal-title":"FEMS Microbiol. Lett."},{"key":"e_1_3_2_19_2","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1111\/j.1365-2958.1994.tb00453.x","volume":"13","year":"1994","unstructured":"Klarsfeld, A. D., P. L. Goossens, and P. Cossart. 1994. Five Listeria monocytogenes genes preferentially expressed in infected mammalian cells: plcA, purH, purD, pyrE and an arginine ABC transporter gene, arpJ. Mol. Microbiol. 13 : 585-597.","journal-title":"Mol. Microbiol."},{"key":"e_1_3_2_20_2","doi-asserted-by":"crossref","first-page":"1207","DOI":"10.1099\/mic.0.2006\/004747-0","volume":"153","year":"2007","unstructured":"Klumpp, J., and T. M. Fuchs. 2007. Identification of novel genes in genomic islands that contribute to Salmonella typhimurium replication in macrophages. Microbiology 153 : 1207-1220.","journal-title":"Microbiology"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1038\/227680a0"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.61.9.3756-3760.1993"},{"key":"e_1_3_2_23_2","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1098\/rstb.2000.0600","volume":"355","year":"2000","unstructured":"Merrell, D. S., and A. Camilli. 2000. Detection and analysis of gene expression during infection by in vivo expression technology. Phil. Trans. R. Soc. Lond. B Biol. Sci. 355 : 587-599.","journal-title":"Phil. Trans. R. Soc. Lond. B Biol. Sci."},{"key":"e_1_3_2_24_2","doi-asserted-by":"crossref","first-page":"790","DOI":"10.1038\/nbt834","volume":"21","year":"2003","unstructured":"Morett, E., J. O. Korbel, E. Rajan, G. Saab-Rincon, L. Olvera, M. Olvera, S. Schmidt, B. Snel, and P. Bork. 2003. Systematic discovery of analogous enzymes in thiamin biosynthesis. Nat. Biotechnol. 21 : 790-795.","journal-title":"Nat. Biotechnol."},{"key":"e_1_3_2_25_2","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1111\/j.1462-5822.2005.00648.x","volume":"8","year":"2006","unstructured":"Munoz-Elias, E. J., and J. D. McKinney. 2006. Carbon metabolism of intracellular bacteria. Cell. Microbiol. 8 : 10-22.","journal-title":"Cell. Microbiol."},{"key":"e_1_3_2_26_2","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1126\/science.1088170","volume":"302","year":"2003","unstructured":"O'Riordan, M., M. A. Moors, and D. A. Portnoy. 2003. Listeria intracellular growth and virulence require host-derived lipoic acid. Science 302 : 462-464.","journal-title":"Science"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.186.5.1571-1573.2004"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.57.10.3046-3048.1991"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01208-08"},{"key":"e_1_3_2_30_2","doi-asserted-by":"crossref","first-page":"48949","DOI":"10.1074\/jbc.M208965200","volume":"277","year":"2002","unstructured":"Rodionov, D. A., A. G. Vitreschak, A. A. Mironov, and M. S. Gelfand. 2002. Comparative genomics of thiamin biosynthesis in procaryotes. New genes and regulatory mechanisms. J. Biol. Chem. 277 : 48949-48959.","journal-title":"J. Biol. Chem."},{"key":"e_1_3_2_31_2","unstructured":"Molecular cloning: a laboratory manual 2001"},{"key":"e_1_3_2_32_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.187.23.8127-8136.2005"},{"key":"e_1_3_2_33_2","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1086\/381206","volume":"189","year":"2004","unstructured":"Slaghuis, J., M. Goetz, F. Engelbrecht, and W. Goebel. 2004. Inefficient replication of Listeria innocua in the cytosol of mammalian cells. J. Infect. Dis. 189 : 393-401.","journal-title":"J. Infect. Dis."},{"key":"e_1_3_2_34_2","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/1471-2091-3-8","volume":"3","year":"2002","unstructured":"Song, Q., and C. K. Singleton. 2002. Mitochondria from cultured cells derived from normal and thiamine-responsive megaloblastic anemia individuals efficiently import thiamine diphosphate. BMC Biochem. 3 : 8.","journal-title":"BMC Biochem."},{"key":"e_1_3_2_35_2","doi-asserted-by":"crossref","first-page":"2319","DOI":"10.1021\/bi0478648","volume":"44","year":"2005","unstructured":"Toms, A. V., A. L. Haas, J. H. Park, T. P. Begley, and S. E. Ealick. 2005. Structural characterization of the regulatory proteins TenA and TenI from Bacillus subtilis and identification of TenA as a thiaminase II. Biochemistry 44 : 2319-2329.","journal-title":"Biochemistry"},{"key":"e_1_3_2_36_2","doi-asserted-by":"crossref","first-page":"8946","DOI":"10.1074\/jbc.273.15.8946","volume":"273","year":"1998","unstructured":"Webb, E., K. Claas, and D. Downs. 1998. thiBPQ encodes an ABC transporter required for transport of thiamine and thiamine pyrophosphate in Salmonella typhimurium. J. Biol. Chem. 273 : 8946-8950.","journal-title":"J. Biol. Chem."},{"key":"e_1_3_2_37_2","doi-asserted-by":"crossref","first-page":"952","DOI":"10.1038\/nature01145","volume":"419","year":"2002","unstructured":"Winkler, W., A. Nahvi, and R. R. Breaker. 2002. Thiamine derivatives bind messenger RNAs directly to regulate bacterial gene expression. Nature 419 : 952-956.","journal-title":"Nature"}],"container-title":["Journal of Bacteriology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JB.01636-08","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JB.01636-08","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,5,23]],"date-time":"2023-05-23T20:40:13Z","timestamp":1684874413000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/JB.01636-08"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2009,4]]},"references-count":36,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2009,4]]}},"alternative-id":["10.1128\/JB.01636-08"],"URL":"https:\/\/doi.org\/10.1128\/jb.01636-08","relation":{},"ISSN":["0021-9193","1098-5530"],"issn-type":[{"value":"0021-9193","type":"print"},{"value":"1098-5530","type":"electronic"}],"subject":[],"published":{"date-parts":[[2009,4]]},"assertion":[{"value":"2008-11-18","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2009-01-20","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-12-20","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}