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Please use this identifier to cite or link to this item: http://repository.iitr.ac.in/handle/123456789/1123
Title: Fosfomycin resistance in Acinetobacter baumannii is mediated by efflux through a major facilitator superfamily (MFS) transporter-AbaF
Authors: Sharma A.
Sharma R.
Bhattacharyya T.
Bhando T.
Pathania, R.
Published in: Journal of Antimicrobial Chemotherapy
Abstract: Objectives: To decipher the function of A1S_1331, named AbaF (Acinetobacter baumannii Fosfomycin efflux), one of the primary targets of AbsR25, a small RNA of A. baumannii. Methods: abaF was cloned in a multicopy plasmid and expressed from its native promoter in an efflux-deficient strain-Escherichia coli KAM32. Drug susceptibility, accumulation and efflux of ethidium bromide (EtBr) were determined in this strain. abaF was disrupted in A. baumannii using homologous recombination and its effect on drug susceptibility, biofilm formation and virulence was studied. Expression of abaF was followed by quantitative PCR in fosfomycin-challenged A. baumannii and fosfomycin-resistant mutants of A. baumannii. Expression of abaF in clinical strains of A. baumannii was determined by RT-PCR. Results: Expression of abaF in E. coli KAM32 resulted in increased resistance to fosfomycin. Lower accumulation and higher efflux of EtBr fromthis strain confirmed the role of AbaFas an efflux pump. Disruption of abaF in A. baumannii caused an increase in fosfomycin susceptibility and a decrease in biofilmformation and virulence. The expression of abaF was higher in A. baumannii cells exposed to fosfomycin and in cells resistant to higher concentrations of fosfomycin. The clinically relevant strains of A. baumannii also tested positive for the expression of abaF. Conclusions: The results of this study suggest that efflux is an important mechanismof fosfomycin resistance and AbaF is involved in fosfomycin resistance in A. baumannii. AbaF also seems to play a role in biofilm formation and virulence of A. baumannii. © The Author 2016.
Citation: Journal of Antimicrobial Chemotherapy (2017), 72(1): 68-74
URI: https://doi.org/10.1093/jac/dkw382
http://repository.iitr.ac.in/handle/123456789/1123
Issue Date: 2017
Publisher: Oxford University Press
ISSN: 3057453
Author Scopus IDs: 57217481823
57212897407
56763662400
56266736600
7004308029
Author Affiliations: Sharma, A., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247667, India
Sharma, R., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247667, India
Bhattacharyya, T., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247667, India
Bhando, T., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247667, India
Pathania, R., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247667, India
Funding Details: We are thankful to Dr Ashima Bhardwaj, IIAR, Gandhinagar, India, for providing E. coli KAM32 as a kind gift and Professor T. Tsuchiya for agreeing to provide E. coli KAM32. We would also like to acknowledge Dr Varsha Gupta, GMCH, Chandigarh, India, for providing the clinical strains of A. baumannii and Dr Soumya Roy Choudhary, IMTECH, Chandigarh, India, for providing E. coli S17-1 lpir. We also thank Dr Naveen K. Navani for his valuable suggestions throughout this study and help in qRT-PCR experiments. We are thankful to Mr Amit Gaurav and Ms Anjul Saini for technical assistance. This work was partially supported by research grant no. AMR/15/2011-ECD-I, Indian Council of Medical Research, New Delhi, India and BT/PR11943/MED/29/874/2014, Department of Biotechnology, Government of India (to R. P.). A. S. and T. Bhando are financially supported by the Ministry of Human Resource Development (MHRD), Government of India.
Corresponding Author: Pathania, R.; Department of Biotechnology, Indian Institute of Technology RoorkeeIndia; email: ranjanapathania@gmail.com
Appears in Collections:Journal Publications [BT]

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