4'-Phosphopantetheinyl transferase PptT, a new drug target required for Mycobacterium tuberculosis growth and persistence in vivo.
The cell envelope of Mycobacterium tuberculosis, the causative agent of tuberculosis in humans, contains lipids with unusual structures. These lipids play a key role in both virulence and resistance to the various hostile environments encountered by the bacteria during infection. They are synthesize...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-12-01
|
| Series: | PLoS Pathogens |
| Online Access: | http://europepmc.org/articles/PMC3534377?pdf=render |
| _version_ | 1811259438517977088 |
|---|---|
| author | Cécile Leblanc Thomas Prudhomme Guillaume Tabouret Aurélie Ray Sophie Burbaud Stéphanie Cabantous Lionel Mourey Christophe Guilhot Christian Chalut |
| author_facet | Cécile Leblanc Thomas Prudhomme Guillaume Tabouret Aurélie Ray Sophie Burbaud Stéphanie Cabantous Lionel Mourey Christophe Guilhot Christian Chalut |
| author_sort | Cécile Leblanc |
| collection | DOAJ |
| description | The cell envelope of Mycobacterium tuberculosis, the causative agent of tuberculosis in humans, contains lipids with unusual structures. These lipids play a key role in both virulence and resistance to the various hostile environments encountered by the bacteria during infection. They are synthesized by complex enzymatic systems, including type-I polyketide synthases and type-I and -II fatty acid synthases, which require a post-translational modification to become active. This modification consists of the covalent attachment of the 4'-phosphopantetheine moiety of Coenzyme A catalyzed by phosphopantetheinyl transferases (PPTases). PptT, one of the two PPTases produced by mycobacteria, is involved in post-translational modification of various type-I polyketide synthases required for the formation of both mycolic acids and lipid virulence factors in mycobacteria. Here we identify PptT as a new target for anti-tuberculosis drugs; we address all the critical issues of target validation to demonstrate that PptT can be used to search for new drugs. We confirm that PptT is essential for the growth of M. bovis BCG in vitro and show that it is required for persistence of M. bovis BCG in both infected macrophages and immunodeficient mice. We generated a conditional expression mutant of M. tuberculosis, in which the expression of the pptT gene is tightly regulated by tetracycline derivatives. We used this construct to demonstrate that PptT is required for the replication and survival of the tubercle bacillus during the acute and chronic phases of infection in mice. Finally, we developed a robust and miniaturized assay based on scintillation proximity assay technology to search for inhibitors of PPTases, and especially of PptT, by high-throughput screening. Our various findings indicate that PptT meets the key criteria for being a therapeutic target for the treatment of mycobacterial infections. |
| first_indexed | 2024-04-12T18:30:38Z |
| format | Article |
| id | doaj.art-e66859b6949c4ac2bb2cb62a24273993 |
| institution | Directory Open Access Journal |
| issn | 1553-7366 1553-7374 |
| language | English |
| last_indexed | 2024-04-12T18:30:38Z |
| publishDate | 2012-12-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj.art-e66859b6949c4ac2bb2cb62a242739932022-12-22T03:21:05ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742012-12-01812e100309710.1371/journal.ppat.10030974'-Phosphopantetheinyl transferase PptT, a new drug target required for Mycobacterium tuberculosis growth and persistence in vivo.Cécile LeblancThomas PrudhommeGuillaume TabouretAurélie RaySophie BurbaudStéphanie CabantousLionel MoureyChristophe GuilhotChristian ChalutThe cell envelope of Mycobacterium tuberculosis, the causative agent of tuberculosis in humans, contains lipids with unusual structures. These lipids play a key role in both virulence and resistance to the various hostile environments encountered by the bacteria during infection. They are synthesized by complex enzymatic systems, including type-I polyketide synthases and type-I and -II fatty acid synthases, which require a post-translational modification to become active. This modification consists of the covalent attachment of the 4'-phosphopantetheine moiety of Coenzyme A catalyzed by phosphopantetheinyl transferases (PPTases). PptT, one of the two PPTases produced by mycobacteria, is involved in post-translational modification of various type-I polyketide synthases required for the formation of both mycolic acids and lipid virulence factors in mycobacteria. Here we identify PptT as a new target for anti-tuberculosis drugs; we address all the critical issues of target validation to demonstrate that PptT can be used to search for new drugs. We confirm that PptT is essential for the growth of M. bovis BCG in vitro and show that it is required for persistence of M. bovis BCG in both infected macrophages and immunodeficient mice. We generated a conditional expression mutant of M. tuberculosis, in which the expression of the pptT gene is tightly regulated by tetracycline derivatives. We used this construct to demonstrate that PptT is required for the replication and survival of the tubercle bacillus during the acute and chronic phases of infection in mice. Finally, we developed a robust and miniaturized assay based on scintillation proximity assay technology to search for inhibitors of PPTases, and especially of PptT, by high-throughput screening. Our various findings indicate that PptT meets the key criteria for being a therapeutic target for the treatment of mycobacterial infections.http://europepmc.org/articles/PMC3534377?pdf=render |
| spellingShingle | Cécile Leblanc Thomas Prudhomme Guillaume Tabouret Aurélie Ray Sophie Burbaud Stéphanie Cabantous Lionel Mourey Christophe Guilhot Christian Chalut 4'-Phosphopantetheinyl transferase PptT, a new drug target required for Mycobacterium tuberculosis growth and persistence in vivo. PLoS Pathogens |
| title | 4'-Phosphopantetheinyl transferase PptT, a new drug target required for Mycobacterium tuberculosis growth and persistence in vivo. |
| title_full | 4'-Phosphopantetheinyl transferase PptT, a new drug target required for Mycobacterium tuberculosis growth and persistence in vivo. |
| title_fullStr | 4'-Phosphopantetheinyl transferase PptT, a new drug target required for Mycobacterium tuberculosis growth and persistence in vivo. |
| title_full_unstemmed | 4'-Phosphopantetheinyl transferase PptT, a new drug target required for Mycobacterium tuberculosis growth and persistence in vivo. |
| title_short | 4'-Phosphopantetheinyl transferase PptT, a new drug target required for Mycobacterium tuberculosis growth and persistence in vivo. |
| title_sort | 4 phosphopantetheinyl transferase pptt a new drug target required for mycobacterium tuberculosis growth and persistence in vivo |
| url | http://europepmc.org/articles/PMC3534377?pdf=render |
| work_keys_str_mv | AT cecileleblanc 4phosphopantetheinyltransferaseppttanewdrugtargetrequiredformycobacteriumtuberculosisgrowthandpersistenceinvivo AT thomasprudhomme 4phosphopantetheinyltransferaseppttanewdrugtargetrequiredformycobacteriumtuberculosisgrowthandpersistenceinvivo AT guillaumetabouret 4phosphopantetheinyltransferaseppttanewdrugtargetrequiredformycobacteriumtuberculosisgrowthandpersistenceinvivo AT aurelieray 4phosphopantetheinyltransferaseppttanewdrugtargetrequiredformycobacteriumtuberculosisgrowthandpersistenceinvivo AT sophieburbaud 4phosphopantetheinyltransferaseppttanewdrugtargetrequiredformycobacteriumtuberculosisgrowthandpersistenceinvivo AT stephaniecabantous 4phosphopantetheinyltransferaseppttanewdrugtargetrequiredformycobacteriumtuberculosisgrowthandpersistenceinvivo AT lionelmourey 4phosphopantetheinyltransferaseppttanewdrugtargetrequiredformycobacteriumtuberculosisgrowthandpersistenceinvivo AT christopheguilhot 4phosphopantetheinyltransferaseppttanewdrugtargetrequiredformycobacteriumtuberculosisgrowthandpersistenceinvivo AT christianchalut 4phosphopantetheinyltransferaseppttanewdrugtargetrequiredformycobacteriumtuberculosisgrowthandpersistenceinvivo |