title: Analysis of uracil phosphoribosyltransferase expression in Mycobacterium tuberculosis and evaluation of upp knockout strain in infected mice.
abstract: The upp (Rv3309c)-encoded uracil phosphoribosyltransferase from Mycobacterium tuberculosis (MtUPRT) converts uracil and 5-phosphoribosyl-α-1-pyrophosphate into pyrophosphate and uridine 5΄-monophosphate, the precursor of all pyrimidine nucleotides. A M. tuberculosis knockout strain for upp gene was generated by allelic replacement. Knockout and complemented strains were validated by a functional assay of uracil incorporation. A basal level of MtUPRT expression is shown to be independent of either growth medium used, addition of bases, or oxygen presence/absence. The upp disruption does not affect M. tuberculosis growth in Middlebrook 7H9 medium, and it is not required for M. tuberculosis virulence in a mouse model of infection. Thus, MtUPRT is unlikely to be a good target for drugs against M. tuberculosis.
authors.affiliation: National Institute of Science and Technology in Tuberculosis, Research Center in Molecular and Functional Biology, Pontifical Catholic University of Rio Grande do Sul-PUCRS, CEP 90619-900, Porto Alegre, RS, Brazil.
authors.forename: Anne Drumond
authors.initials: AD
authors.lastname: Villela
authors.affiliation: Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523 USA.
authors.forename: Ha
authors.initials: H
authors.lastname: Pham
authors.affiliation: Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523 USA.
authors.forename: Victoria
authors.initials: V
authors.lastname: Jones
authors.affiliation: Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523 USA.
authors.forename: Anna E
authors.initials: AE
authors.lastname: Grzegorzewicz
authors.affiliation: National Institute of Science and Technology in Tuberculosis, Research Center in Molecular and Functional Biology, Pontifical Catholic University of Rio Grande do Sul-PUCRS, CEP 90619-900, Porto Alegre, RS, Brazil.|Graduate Program in Medicine and Health Sciences, PUCRS, CEP 90610-000, Porto Alegre, Brazil.
authors.forename: Valnês da Silva
authors.initials: VD
authors.lastname: Rodrigues-Junior
authors.affiliation: National Institute of Science and Technology in Tuberculosis, Research Center in Molecular and Functional Biology, Pontifical Catholic University of Rio Grande do Sul-PUCRS, CEP 90619-900, Porto Alegre, RS, Brazil.|Graduate Program in Medicine and Health Sciences, PUCRS, CEP 90610-000, Porto Alegre, Brazil.
authors.forename: Maria Martha
authors.initials: MM
authors.lastname: Campos
authors.affiliation: National Institute of Science and Technology in Tuberculosis, Research Center in Molecular and Functional Biology, Pontifical Catholic University of Rio Grande do Sul-PUCRS, CEP 90619-900, Porto Alegre, RS, Brazil.|Graduate Program in Medicine and Health Sciences, PUCRS, CEP 90610-000, Porto Alegre, Brazil.|Graduate Program in Celular and Molecular Biology, PUCRS, CEP 90619-900, Porto Alegre, Brazil.
authors.forename: Luiz Augusto
authors.initials: LA
authors.lastname: Basso
authors.affiliation: Graduate Program in Celular and Molecular Biology, PUCRS, CEP 90619-900, Porto Alegre, Brazil.
authors.forename: Mary
authors.initials: M
authors.lastname: Jackson
authors.affiliation: National Institute of Science and Technology in Tuberculosis, Research Center in Molecular and Functional Biology, Pontifical Catholic University of Rio Grande do Sul-PUCRS, CEP 90619-900, Porto Alegre, RS, Brazil.|Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523 USA.
authors.forename: Diógenes Santiago
authors.initials: DS
authors.lastname: Santos
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D015870:Gene Expression; D055786:Gene Knockout Techniques; D051379:Mice; D009169:Mycobacterium tuberculosis / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D010430:Pentosyltransferases / Q000235:genetics* / Q000378:metabolism; D014376:Tuberculosis / Q000382:microbiology*; D014498:Uracil / Q000378:metabolism / Q000494:pharmacology; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D014498:Uracil; D010430:Pentosyltransferases; C023269:uracil phosphoribosyltransferase
keywords: Mycobacterium tuberculosis; Wayne model; gene knockout; mouse model; upp gene; uracil phosphoribosyltransferase