Delineation of the Residues of Bacillus anthracis Zinc Uptake Regulator Protein Directly Involved in Its Interaction wit
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Delineation of the Residues of Bacillus anthracis Zinc Uptake Regulator Protein Directly Involved in Its Interaction with Cognate DNA Divya Kandari 1 & Hemant Joshi 1 & Neetu Tanwar 2 & Manoj Munde 2 & Rakesh Bhatnagar 1,3 Received: 12 July 2020 / Accepted: 5 October 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Zinc uptake regulator (Zur) is a negative transcriptional regulator of bacteria that belongs to the FUR superfamily of proteins and regulates zinc (Zn) homeostasis under extreme Zn conditions. The Zur protein of Bacillus anthracis (BaZur) was though characterized previously, but the residues of this transcriptional regulator, crucial for binding to the consensus Zur box in the cognate DNA, remain unexplored. In this study, we reveal the essential residues of the protein that govern the specific interaction with the cognate DNA, through mutational and binding studies. In silico predicted model of the BaZur protein with the promoter region of one of the regulon candidates was utilized to identify specific residues of the N-terminal domain (NTD), constituting the DNA-binding recognition helix. Our results suggest that two phenylalanine residues, a non-polar aliphatic leucine and a positively charged arginine residue of NTD, are predominantly involved in DNA binding of BaZur. Among these, the arginine residue (Arg58) is conserved among all the Zur proteins and the two Phe residues, namely Phe53 and Phe63, are conserved in the Zur proteins of Staphylococcus aureus and Listeria monocytogenes. Taken together, the current study represents an in-depth investigation into the key DNA-binding residues involved in the BaZur-DNA interaction. Keywords Bacillus anthracis . Zinc uptake regulator (Zur) . Transcriptional factor . DNA-protein interaction . Zn homeostasis
Introduction Post invasion pathogens continuously struggle to thrive under the tremendous pressure of nutritional immunity imposed by the host immune system. Nutritional immunity is the intrinsic defence mechanism wherein the host either starves or flushes excess of crucial metal ions onto the pathogen [16, 34]. One such crucial transition metal ion for all living systems is Zn. It
Divya Kandari and Hemant Joshi contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12011-020-02427-x) contains supplementary material, which is available to authorized users. * Rakesh Bhatnagar [email protected] 1
Molecular Biology and Genetic Engineering Laboratory, School of Biotechnology, Jawaharlal Nehru University, New Delhi 110067, India
2
School of Physical Sciences, Jawaharlal Nehru University, New Delhi 110067, India
3
Banaras Hindu University, Banaras, Uttar Pradesh 221005, India
is needed for a gamut of functions inclusive of regulatory, catalytic, and structural [3]. Also, cardinal cellular processes like DNA proliferation, DNA damage repair, and cell division are Zn-dependent [1, 15]. Despite the pivotal roles played by Zn in general cellula
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