Abstract:
:Oligopeptides play important roles in bacterial nutrition and signaling. Using sequences from the available genome database for Mycobacterium tuberculosis H37Rv, the oligopeptide permease operon (oppBCDA) of Mycobacterium bovis BCG was cloned from a cosmid library. An opp mutant strain was constructed by homologous recombination with an allele of oppD interrupted by kanamycin and streptomycin resistance markers. The deletion was complemented with a wild-type copy of the opp operon. Two approaches were taken to characterize the peptide transporter defect in this mutant strain. First, growth of wild-type and mutant strains was monitored in media containing a wide variety of peptides as sole source of carbon and/or nitrogen. Among 25 peptides ranging from two to six amino acids in length, none was capable of supporting measurable growth as the sole carbon source in either wild-type or mutant strains. The second approach exploited the resistance of permease mutants to toxic substrates. The tripeptide glutathione (gamma-glutamyl-L-cyteinylglycine [GSH]) is toxic to wild-type BCG and was used successfully to characterize peptide uptake in the opp mutant. In 2 mM GSH, growth of the wild-type strain is inhibited, whereas the opp mutant is resistant to concentrations as high as 10 mM. Similar results were found with the tripeptide S-nitrosoglutathione (GSNO), thought to be a donor of NO in mammalian cells. Using incorporation of [(3)H]uracil to monitor the effects of GSH and GSNO on macromolecular synthesis in growing cells, it was demonstrated that the opp mutant is resistant, whereas the wild type and the mutant complemented with a wild-type copy of the operon are sensitive to both tripeptides. In uptake measurements, incorporation of [(3)H]GSH is reduced in the mutant compared with wild type and the complemented mutant. Finally, growth of the three strains in the tripeptides suggests that GSH is bacteriostatic, whereas GSNO is bacteriocidal.
journal_name
Infect Immunjournal_title
Infection and immunityauthors
Green RM,Seth A,Connell NDdoi
10.1128/iai.68.2.429-436.2000keywords:
subject
Has Abstractpub_date
2000-02-01 00:00:00pages
429-36issue
2eissn
0019-9567issn
1098-5522journal_volume
68pub_type
杂志文章abstract::We have determined the nucleotide sequence of the 1.4-kilobase DNA fragment containing the plasminogen activator gene (pla) of Yersinia pestis, which determines both plasminogen activator and coagulase activities of the species. The sequence revealed the presence of a 936-base-pair open reading frame that constitutes ...
journal_title:Infection and immunity
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doi:10.1128/IAI.57.5.1517-1523.1989
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journal_title:Infection and immunity
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.1.6.521-525.1970
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journal_title:Infection and immunity
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journal_title:Infection and immunity
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doi:10.1128/IAI.22.2.359-364.1978
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journal_title:Infection and immunity
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journal_title:Infection and immunity
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更新日期:1974-05-01 00:00:00
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journal_title:Infection and immunity
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doi:10.1128/IAI.63.3.961-968.1995
更新日期:1995-03-01 00:00:00
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journal_title:Infection and immunity
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journal_title:Infection and immunity
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更新日期:1976-04-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.43.2.730-734.1984
更新日期:1984-02-01 00:00:00