Transposon Mutagenesis in Mycobacterium avium Subspecies Paratuberculosis

Bannantine, John P.; Zinniel, Denise K.; Barletta, Raúl G.

doi:10.1007/978-1-4939-9570-7_11

Cited by 2 publications

(4 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“… 40 , 79 , 80 TMM is essential for mycobacteria cell viability, and therefore many proteins involved in the synthesis of TMM are also essential. 81–85 Consistent with a model of essentiality, enzymes involved in MA synthesis are often identified as targets of anti-TB inhibitors. Notable targets include InhA and HadAB, which are inhibited by the TB drugs INH 56 /ethionamide 86 , 87 and isoxyl 88 /thiacetazone, 88 respectively.…”

Section: Synthesis and Transport Of Tmm And Tdmmentioning

confidence: 97%

“… 116 However, only mmpL3 and mmpL11 are conserved in all species of mycobacteria 116 and only mmpL3 is essential for growth and viability both in vitro and in vivo in most mycobacteria. 29 , 81–85 , 89 , 135–140 …”

Section: Mmpl3 Protein Structure and Functionmentioning

confidence: 99%

“… 137 , 138 , 140 In addition, saturating transposon mutagenesis studies have failed to identify null mutants in Mtb, Mycobacterium bovis Bacillus Calmette-Guérin (BCG), or Mycobacterium paratuberculosis . 81–85 , 135 However, recently, Xiong et al reported an mmpL3 knockout in M. neoaurum (ATCC 25795). 139 The method used to create this knockout strain failed to generate knockouts in Mtb 143 and M. smegmatis .…”

Section: The Therapeutic Potential Of Mmpl3mentioning

confidence: 99%

See 2 more Smart Citations

Molecular Mechanisms of MmpL3 Function and Inhibition

Williams

Abramovitch

2023

Microbial Drug Resistance

View full text Add to dashboard Cite

Mycobacteria species include a large number of pathogenic organisms such as Mycobacterium tuberculosis , Mycobacterium leprae , and various non-tuberculous mycobacteria. Mycobacterial membrane protein large 3 (MmpL3) is an essential mycolic acid and lipid transporter required for growth and cell viability. In the last decade, numerous studies have characterized MmpL3 with respect to protein function, localization, regulation, and substrate/inhibitor interactions. This review summarizes new findings in the field and seeks to assess future areas of research in our rapidly expanding understanding of MmpL3 as a drug target. An atlas of known MmpL3 mutations that provide resistance to inhibitors is presented, which maps amino acid substitutions to specific structural domains of MmpL3. In addition, chemical features of distinct classes of Mmpl3 inhibitors are compared to provide insights into shared and unique features of varied MmpL3 inhibitors.

show abstract

Section: Synthesis and Transport Of Tmm And Tdmmentioning

confidence: 97%

Section: Mmpl3 Protein Structure and Functionmentioning

confidence: 99%

Section: The Therapeutic Potential Of Mmpl3mentioning

confidence: 99%

See 1 more Smart Citation

Molecular Mechanisms of MmpL3 Function and Inhibition

Williams

Abramovitch

2023

Microbial Drug Resistance

View full text Add to dashboard Cite

show abstract

“…Previous research on mycobacterial pathogens has led to the consensus that live-attenuated vaccines are the best approach for vaccination [ 24 ]. Our laboratory has established a large MAP collection comprising more than a million mutants, including Tn 5367 and Himar1 transposon insertion mutant banks [ 25 , 26 , 27 ]. In addition, two deletion mutants (DMAP52 and DMAP56) in ORFs MAP_1152 and MAP_1156 were isolated.…”

Section: Introductionmentioning

confidence: 99%

Mycobacterium avium subsp. paratuberculosis Candidate Vaccine Strains Are Pro-apoptotic in RAW 264.7 Murine Macrophages

et al. 2023

Self Cite

View full text Add to dashboard Cite

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of Johne’s disease, a severe gastroenteritis of ruminants. This study developed a model cell culture system to rapidly screen MAP mutants with vaccine potential for apoptosis. Two wild-type strains, a transposon mutant, and two deletion mutant MAP strains (MOI of 10 with 1.2 × 106 CFU) were tested in murine RAW 264.7 macrophages to determine if they induce apoptosis and/or necrosis. Both deletion mutants were previously shown to be attenuated and immunogenic in primary bovine macrophages. All strains had similar growth rates, but cell morphology indicated that both deletion mutants were elongated with cell wall bulging. Cell death kinetics were followed by a real-time cellular assay to measure luminescence (apoptosis) and fluorescence (necrosis). A 6 h infection period was the appropriate time to assess apoptosis that was followed by secondary necrosis. Apoptosis was also quantified via DAPI-stained nuclear morphology and validated via flow cytometry. The combined analysis confirmed the hypothesis that candidate vaccine deletion mutants are pro-apoptotic in RAW 264.7 cells. In conclusion, the increased apoptosis seen in the deletion mutants correlates with the attenuated phenotype and immunogenicity observed in bovine macrophages, a property associated with good vaccine candidates.

show abstract

Transposon Mutagenesis in Mycobacterium avium Subspecies Paratuberculosis

Cited by 2 publications

References 22 publications

Molecular Mechanisms of MmpL3 Function and Inhibition

Molecular Mechanisms of MmpL3 Function and Inhibition

Mycobacterium avium subsp. paratuberculosis Candidate Vaccine Strains Are Pro-apoptotic in RAW 264.7 Murine Macrophages

Contact Info

Product

Resources

About