TtgB Notes
Gene Overview
TtgB (PP_1385, Q88N31) is the inner membrane RND (Resistance-Nodulation-Division) transporter component of the TtgABC tripartite efflux pump in Pseudomonas putida KT2440. It is a 1050 AA protein with 12 predicted transmembrane helices, belonging to the acriflavine resistance protein family (IPR001036) and specifically the MdtF-like subfamily (IPR004764). It is homologous to AcrB in E. coli and MexB in P. aeruginosa.
TtgABC System Architecture
The TtgABC efflux pump is a tripartite system typical of RND family transporters:
- TtgA (PP_1384): Membrane fusion protein (MFP/periplasmic adaptor)
- TtgB (PP_1385): Inner membrane RND transporter (proton-dependent substrate extrusion)
- TtgC (PP_1386): Outer membrane factor (OMF/channel)
The three genes form an operon, transcribed from a single promoter upstream of ttgA. Expression is constitutive but regulated by the TtgR repressor (PP_1387), which is transcribed divergently from ttgA PMID:11251828.
Substrate Specificity and Function
Organic Solvents
In DOT-T1E strain, TtgABC extrudes toluene, styrene, m-xylene, ethylbenzene, and propylbenzene PMID:11395460. However, in KT2440 this operon may not function primarily in toluene efflux [UniProt CAUTION: "In KT2400 this operon does not seem to function in toluene efflux"].
Antibiotics
TtgABC confers resistance to multiple antibiotics in KT2440:
- Chloramphenicol: TtgABC knockout is compromised PMID:22143519
- Fluoroquinolones and beta-lactams: Overexpression of TtgABC via TtgR mutations leads to increased resistance PMID:32840000
- Also quinolones, sulfonamides PMID:24907323
- Indole induces ttgABC expression contributing to ampicillin resistance PMID:28352264
Plant Antimicrobials and Bipyridyls
- TtgR binds phloretin and other plant antimicrobials, releasing TtgABC expression PMID:17466326
- TtgABC is required for detoxification of bipyridyl compounds (e.g., caerulomycin A) PMID:32973714
- Also involved in resistance to bile salts and deoxycholate PMID:32973714
Multidrug Binding
TtgB itself has been shown to possess multidrug-binding capacity PMID:17498746.
Additional Roles Beyond Efflux
Knockout of ECF-10 sigma factor leads to upregulation of ttgABC and enhanced biofilm formation, suggesting TtgABC plays a role in biofilm development PMID:24907323.
Ion Homeostasis
In the absence of TtgABC, bipyridyl compounds interfere with metal ion homeostasis, affecting copper availability and pyoverdine (siderophore) production PMID:32973714.
Regulatory Context
- TtgR: Adjacent repressor, binds pseudo-palindromic operator overlapping ttgR/ttgA promoters. Released by effectors including antibiotics and plant antimicrobials [PMID:11251828, PMID:17466326, PMID:20435893]
- ECF-10 (PP4553): Knockout leads to ttgABC upregulation PMID:24907323
- PpeRS: Two-component system positively controls TtgABC expression PMID:32840000
- Lrp-like: Global regulator involved in solvent-tolerant response PMID:11251828
Key Distinction: KT2440 vs DOT-T1E
The UniProt entry is for KT2440. The caution note indicates that while the homologous system in DOT-T1E functions in solvent efflux, in KT2440 it primarily functions in antibiotic resistance, bipyridyl detoxification, and potentially biofilm formation. The proteins are highly similar but expression levels and ecological context differ.
Homologous Systems
- MepABC (P. putida KT2442-TOL): Solvent and antibiotic efflux
- ArpABC (P. putida S12): Antibiotic efflux only, not solvent tolerance PMID:11160799
- SrpABC (P. putida B6-2): PAH degradation support PMID:29030440
- AcrAB-TolC (E. coli): Canonical RND efflux system
- MexAB-OprM (P. aeruginosa): Major multidrug efflux
Energy Coupling
Proton-dependent, operates via proton motive force. Energy transfer may involve the TonB system PMID:12142492.
References
- PMID:9642183 - Original identification of TtgABC efflux pump in DOT-T1E (Ramos et al. 1998)
- PMID:10648517 - Second efflux system TtgDEF (Mosqueda & Ramos 2000)
- PMID:11160799 - ArpABC in S12 multidrug resistance (Kieboom & de Bont 2001)
- PMID:11251828 - TtgR regulation of TtgABC (Duque et al. 2001)
- PMID:11395460 - Three efflux pumps for toluene tolerance (Rojas et al. 2001)
- PMID:12142492 - Review of solvent tolerance mechanisms (Ramos et al. 2002)
- PMID:12534463 - KT2440 genome sequence (Nelson et al. 2002)
- PMID:12743835 - Comparative genomics of solvent extrusion pumps (Segura et al. 2003)
- PMID:16109935 - Proteomics of toluene response (Segura et al. 2005)
- PMID:17466326 - TtgR crystal structures with antibiotics/plant antimicrobials (Alguel et al. 2007)
- PMID:17498746 - TtgR operator binding cooperativity (Krell et al. 2007)
- PMID:17986203 - Cross-regulation of TtgV and TtgT (Teran et al. 2007)
- PMID:20435893 - TtgR domain cross-talk (Daniels et al. 2010)
- PMID:22143519 - Chloramphenicol resistance in KT2440 (Fernandez et al. 2011)
- PMID:24907323 - ECF-10 affects TtgABC, biofilm formation (Tettmann et al. 2014)
- PMID:28352264 - Indole-induced TtgABC and ampicillin resistance (Kim et al. 2017)
- PMID:29030440 - SrpABC/TtgABC in PAH degradation (Yao et al. 2017)
- PMID:32840000 - ParXY/TtgABC multidrug resistance (Puja et al. 2020)
- PMID:32973714 - TtgABC bipyridyl resistance in KT2440 (Henriquez et al. 2020)