Skip to main content
SpringerLink
Log in

Phenotype microarray analysis of the AdeRS two-component system in Acinetobacter baumannii

  • Original Article
  • Published:
European Journal of Clinical Microbiology & Infectious Diseases Aims and scope Submit manuscript

Abstract

Acinetobacter baumannii is a nosocomial pathogen capable of resistance to multiple antimicrobials. The AdeRS two-component system (TCS) is associated with antimicrobial resistance by controlling the AdeABC efflux pump. To elucidate modulation by AdeRS, we made an A. baumannii mutant lacking the AdeRS TCS and characterized it using phenotype microarray (PM) analysis. After disrupting the adeRS operon, lower expression of AdeABC efflux pump was observed in the mutant strain. PM analysis showed that the AdeRS deletion strain and parental strain presented different tolerances to 91 compounds. Tolerance to 54 of the 91 compounds could be restored by complementing the AdeRS deleted strain with a plasmid carrying the adeRS gene. Compared to the parental strain, the AdeRS deletion strain was more sensitive to various inhibitors that target on-protein synthesis and function of cell membrane permeability. Tolerance to phleomycin of the AdeRS deletion strain reduced greatly and was further confirmed with minimum inhibitory concentration (MIC) determination and spot assay. The efflux pump inhibitor, NMP, could reduce phleomycin MIC four-fold at least for 29 (34.8%) of 81 tigecycline-resistant extensively drug-resistant A. baumannii (TGC-resistant XDRAB) clinical isolates. Our results suggested that the AdeRS TCS of A. baumannii was important for both elimination of antibiotics and tolerance to particular compounds.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Gonzalez-Villoria AM, Valverde-Garduno V (2016) Antibiotic-resistant Acinetobacter baumannii increasing success remains a challenge as a nosocomial pathogen. J Pathog 2016:7318075

    Article  PubMed  PubMed Central  Google Scholar 

  2. Deng M, Zhu MH, Li JJ, Bi S, Sheng ZK, Hu FS, Zhang JJ, Chen W, Xue XW, Sheng JF, Li LJ (2014) Molecular epidemiology and mechanisms of tigecycline resistance in clinical isolates of Acinetobacter baumannii from a Chinese university hospital. Antimicrob Agents Chemother 58(1):297–303

    Article  PubMed  PubMed Central  Google Scholar 

  3. Chan MC, Chiu SK, Hsueh PR, Wang NC, Wang CC, Fang CT (2014) Risk factors for healthcare-associated extensively drug-resistant Acinetobacter baumannii infections: a case–control study. PLoS One 9(1):e85973

    Article  PubMed  PubMed Central  Google Scholar 

  4. Sun J, Deng Z, Yan A (2014) Bacterial multidrug efflux pumps: mechanisms, physiology and pharmacological exploitations. Biochem Biophys Res Commun 453(2):254–267

    Article  CAS  PubMed  Google Scholar 

  5. Bernal P, Molina-Santiago C, Daddaoua A, Llamas MA (2013) Antibiotic adjuvants: identification and clinical use. Microb Biotechnol 6(5):445–449

    Article  PubMed  PubMed Central  Google Scholar 

  6. Rumbo C, Gato E, López M, Ruiz de Alegría C, Fernández-Cuenca F, Martínez-Martínez L, Vila J, Pachón J, Cisneros JM, Rodríguez-Baño J, Pascual A, Bou G, Tomás M; Spanish Group of Nosocomial Infections and Mechanisms of Action and Resistance to Antimicrobials (GEIH-GEMARA); Spanish Society of Clinical Microbiology and Infectious Diseases (SEIMC); Spanish Network for Research in Infectious Diseases (REIPI) (2013) Contribution of efflux pumps, porins, and beta-lactamases to multidrug resistance in clinical isolates of Acinetobacter baumannii. Antimicrob Agents Chemother 57(11):5247–5257

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Yoon EJ, Balloy V, Fiette L, Chignard M, Courvalin P, Grillot-Courvalin C (2016) Contribution of the Ade resistance-nodulation-cell division-type efflux pumps to fitness and pathogenesis of Acinetobacter baumannii. MBio 7(3). pii: e00697-16

  8. Sun JR, Perng CL, Lin JC, Yang YS, Chan MC, Chang TY, Lin FM, Chiueh TS (2014) AdeRS combination codes differentiate the response to efflux pump inhibitors in tigecycline-resistant isolates of extensively drug-resistant Acinetobacter baumannii. Eur J Clin Microbiol Infect Dis 33(12):2141–2147

    Article  CAS  PubMed  Google Scholar 

  9. Sun JR, Perng CL, Chan MC, Morita Y, Lin JC, Su CM, Wang WY, Chang TY, Chiueh TS (2012) A truncated AdeS kinase protein generated by ISAba1 insertion correlates with tigecycline resistance in Acinetobacter baumannii. PLoS One 7(11):e49534

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Sun JR, Jeng WY, Perng CL, Yang YS, Soo PC, Chiang YS, Chiueh TS (2016) Single amino acid substitution Gly186Val in AdeS restores tigecycline susceptibility of Acinetobacter baumannii. J Antimicrob Chemother 71(6):1488–1492

    Article  CAS  PubMed  Google Scholar 

  11. Lin MF, Lin YY, Lan CY (2015) The role of the two-component system BaeSR in disposing chemicals through regulating transporter systems in Acinetobacter baumannii. PLoS One 10(7):e0132843

    Article  PubMed  PubMed Central  Google Scholar 

  12. Richmond GE, Evans LP, Anderson MJ, Wand ME, Bonney LC, Ivens A, Chua KL, Webber MA, Sutton JM, Peterson ML, Piddock LJ (2016) The Acinetobacter baumannii two-component system AdeRS regulates genes required for multidrug efflux, biofilm formation, and virulence in a strain-specific manner. MBio 7(2):e00430-16

    Article  PubMed  PubMed Central  Google Scholar 

  13. Szurmant H, White RA, Hoch JA (2007) Sensor complexes regulating two-component signal transduction. Curr Opin Struct Biol 17(6):706–715

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Balasubramanian D, Schneper L, Kumari H, Mathee K (2013) A dynamic and intricate regulatory network determines Pseudomonas aeruginosa virulence. Nucleic Acids Res 41(1):1–20

    Article  CAS  PubMed  Google Scholar 

  15. Gooderham WJ, Hancock RE (2009) Regulation of virulence and antibiotic resistance by two-component regulatory systems in Pseudomonas aeruginosa. FEMS Microbiol Rev 33(2):279–294

    Article  CAS  PubMed  Google Scholar 

  16. Kröger C, Kary SC, Schauer K, Cameron AD (2016) Genetic regulation of virulence and antibiotic resistance in Acinetobacter baumannii. Genes (Basel) 8(1). pii: E12

  17. Yoon EJ, Chabane YN, Goussard S, Snesrud E, Courvalin P, Dé E, Grillot-Courvalin C (2015) Contribution of resistance-nodulation-cell division efflux systems to antibiotic resistance and biofilm formation in Acinetobacter baumannii. MBio 6(2). pii: e00309-15

  18. Zhang J, Biswas I (2009) A phenotypic microarray analysis of a Streptococcus mutans liaS mutant. Microbiology 155(Pt 1):61–68

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Li L, Hassan KA, Brown MH, Paulsen IT (2016) Rapid multiplexed phenotypic screening identifies drug resistance functions for three novel efflux pumps in Acinetobacter baumannii. J Antimicrob Chemother 71(5):1223–1232

    Article  PubMed  Google Scholar 

  20. Mishra MN, Daniels L (2013) Characterization of the MSMEG_2631 gene (mmp) encoding a multidrug and toxic compound extrusion (MATE) family protein in Mycobacterium smegmatis and exploration of its polyspecific nature using biolog phenotype microarray. J Bacteriol 195(7):1610–1621

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Sun JR, Chan MC, Chang TY, Wang WY, Chiueh TS (2010) Overexpression of the adeB gene in clinical isolates of tigecycline-nonsusceptible Acinetobacter baumannii without insertion mutations in adeRS. Antimicrob Agents Chemother 54(11):4934–4938

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Marchand I, Damier-Piolle L, Courvalin P, Lambert T (2004) Expression of the RND-type efflux pump AdeABC in Acinetobacter baumannii is regulated by the AdeRS two-component system. Antimicrob Agents Chemother 48(9):3298–3304

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Yoon EJ, Courvalin P, Grillot-Courvalin C (2013) RND-type efflux pumps in multidrug-resistant clinical isolates of Acinetobacter baumannii: major role for AdeABC overexpression and AdeRS mutations. Antimicrob Agents Chemother 57(7):2989–2995

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Oh MH, Lee JC, Kim J, Choi CH, Han K (2015) Simple method for markerless gene deletion in multidrug-resistant Acinetobacter baumannii. Appl Environ Microbiol 81(10):3357–3368

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Amin IM, Richmond GE, Sen P, Koh TH, Piddock LJ, Chua KL (2013) A method for generating marker-less gene deletions in multidrug-resistant Acinetobacter baumannii. BMC Microbiol 13:158

    Article  PubMed  PubMed Central  Google Scholar 

  26. Vila J, Martí S, Sánchez-Céspedes J (2007) Porins, efflux pumps and multidrug resistance in Acinetobacter baumannii. J Antimicrob Chemother 59(6):1210–1215

    Article  CAS  PubMed  Google Scholar 

  27. Nikaido H (1989) Outer membrane barrier as a mechanism of antimicrobial resistance. Antimicrob Agents Chemother 33(11):1831–1836

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Cabral JP (1991) Damage to the cytoplasmic membrane and cell death caused by dodine (dodecylguanidine monoacetate) in Pseudomonas syringae ATCC 12271. Antimicrob Agents Chemother 35(2):341–344

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Seo J, Darwin AJ (2013) The Pseudomonas aeruginosa periplasmic protease CtpA can affect systems that impact its ability to mount both acute and chronic infections. Infect Immun 81(12):4561–4570

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Soboh F, Khoury AE, Zamboni AC, Davidson D, Mittelman MW (1995) Effects of ciprofloxacin and protamine sulfate combinations against catheter-associated Pseudomonas aeruginosa biofilms. Antimicrob Agents Chemother 39(6):1281–1286

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Girardello R, Visconde M, Cayô R, Figueiredo RC, Mori MA, Lincopan N, Gales AC (2017) Diversity of polymyxin resistance mechanisms among Acinetobacter baumannii clinical isolates. Diagn Microbiol Infect Dis 87(1):37–44

    Article  CAS  PubMed  Google Scholar 

  32. Olaitan AO, Morand S, Rolain JM (2014) Mechanisms of polymyxin resistance: acquired and intrinsic resistance in bacteria. Front Microbiol 5:643

    Article  PubMed  PubMed Central  Google Scholar 

  33. He X, Lu F, Yuan F, Jiang D, Zhao P, Zhu J, Cheng H, Cao J, Lu G (2015) Biofilm formation caused by clinical Acinetobacter baumannii isolates is associated with overexpression of the AdeFGH efflux pump. Antimicrob Agents Chemother 59(8):4817–4825

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Chen H, Cao J, Zhou C, Liu H, Zhang X, Zhou T (2017) Biofilm formation restrained by subinhibitory concentrations of tigecyclin in Acinetobacter baumannii is associated with downregulation of efflux pumps. Chemotherapy 62(2):128–133

    Article  CAS  PubMed  Google Scholar 

  35. Chankova SG, Dimova E, Dimitrova M, Bryant PE (2007) Induction of DNA double-strand breaks by zeocin in Chlamydomonas reinhardtii and the role of increased DNA double-strand breaks rejoining in the formation of an adaptive response. Radiat Environ Biophys 46(4):409–416

    Article  CAS  PubMed  Google Scholar 

  36. Inaba M, Matsuda N, Banno H, Jin W, Wachino JI, Yamada K, Kimura K, Arakawa Y (2016) In vitro reduction of antibacterial activity of tigecycline against multidrug-resistant Acinetobacter baumannii with host stress hormone norepinephrine. Int J Antimicrob Agents 48(6):680–689

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors thank Ching-Mei Yu (Division of Clinical Pathology, Tri-Service General Hospital) and Ming-Chin Chan (Infection Control Office, Tri-Service General Hospital) for the collection of clinical isolates.

Author information

Authors and Affiliations

  1. Division of Clinical Pathology, Department of Pathology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan

    J.-R. Sun, H.-S. Shang & C.-L. Perng

  2. Graduate Institute of Pathology, National Defense Medical Center, Taipei, Taiwan

    Y.-S. Chiang & C.-L. Perng

  3. Department of Laboratory Medicine, Huilai Medical Care Corporation Hongren Hospital, Yuanlin Township, Changhua County, Taiwan

    Y.-S. Chiang

  4. Division of Infectious Diseases and Tropical Medicine, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan

    Y.-S. Yang

  5. Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, 5 Fu-Hsing Street, Kuei-Shan District, Taoyuan City, Taiwan, Republic of China

    T.-S. Chiueh

Authors
  1. J.-R. Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Y.-S. Chiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. H.-S. Shang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. C.-L. Perng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Y.-S. Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. T.-S. Chiueh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to T.-S. Chiueh.

Ethics declarations

Funding

This study was supported by grants from the Tri-Service General Hospital of the National Defense Medical Center (TSGH-C105-134 and TSGH-C106-117) and the Ministry of Science and Technology (MOST 104-2314-B-016-051 and MOST 105-2628-B-016-003-MY2).

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

Informed consent was obtained from all mothers or legal guardians. This article does not contain any studies with animals performed by any of the authors.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sun, JR., Chiang, YS., Shang, HS. et al. Phenotype microarray analysis of the AdeRS two-component system in Acinetobacter baumannii . Eur J Clin Microbiol Infect Dis 36, 2343–2353 (2017). https://doi.org/10.1007/s10096-017-3066-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10096-017-3066-9

Keywords

Search

Navigation