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Structural Insight into the Gene Expression Profiling of the hcn Operon in Pseudomonas aeruginosa

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Abstract

Pseudomonas aeruginosa is a common opportunistic human pathogen. It generally attacks immunosuppressed patients like AIDS, cancer, cystic fibrosis, etc. The virulence of P. aeruginosa is mediated by various virulence factors. One of such potential virulence factors is HCN synthesized by HCN synthase enzyme, which is encoded by the hcnABC operon. The expressions of the genes of this operon are regulated by three transcriptional regulators, viz., LasR, ANR, and RhlR. In our previous work, we analyzed the molecular details of the functionalities of LasR. In this work, we focused on ANR. ANR is a regulatory protein which belongs to the FNR family and works in anaerobic condition. ANR binds to the promoter DNA, named ANR box, as a dimer. The dimerization of this ANR protein is regulated by Fe4S4, an iron-sulfur cluster. This dimer of ANR (ANR-Fe4S4/ANR-Fe4S4) recognizes and binds the promoter DNA sequence and regulates the transcription of this hcnABC operon. Till date, the biomolecular details of the interactions of ANR dimer with the promoter DNA are not fully understood. Thus, we built the molecular model of ANR-Fe4S4/ANR-Fe4S4. We docked the complex with the corresponding promoter DNA region. We analyzed the mode of interactions with the promoter DNA under different conditions. Thus, we tried to analyze the functionality of the ANR protein during the expressions of the genes of the hcnABC operon. So far, this is the first report to detail the molecular mechanism of the gene expression in P. aeruginosa.

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Acknowledgements

The authors would like to thank the anonymous reviewers for their constructive comments for the betterment of the manuscript. The authors are grateful to the BIF Center, University of Kalyani, for providing the necessary equipment and workstation to carry out the experiments. The authors would like to thank the Department of Biotechnology (DBT, India) for the financial support (SAN No. 102/IFD/ SAN/1824/2015-2016). The infrastructural support from University of Kalyani is also duly acknowledged here. NC would like to acknowledge the UGC, India, for providing the Rajib Gandhi National Fellowship.

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Authors and Affiliations

  1. Department of Biochemistry and Biophysics, University of Kalyani, Kalyani, Nadia, 741235, India

    Nilkanta Chowdhury & Angshuman Bagchi

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  1. Nilkanta Chowdhury
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  2. Angshuman Bagchi
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Correspondence to Angshuman Bagchi.

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The authors declare that they have no conflict of interest.

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Highlights

• Regulations of hcn operon

• Molecular modeling of ANR protein

• Docking of ANR-Fe4S4/ANR-Fe4S4 with promoter DNA

• Molecular mechanism of gene expression

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Chowdhury, N., Bagchi, A. Structural Insight into the Gene Expression Profiling of the hcn Operon in Pseudomonas aeruginosa . Appl Biochem Biotechnol 182, 1144–1157 (2017). https://doi.org/10.1007/s12010-016-2388-x

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  • DOI: https://doi.org/10.1007/s12010-016-2388-x

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