Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2019 Jul;120(7):11318-11330.
doi: 10.1002/jcb.28408. Epub 2019 Feb 5.

Computational and mutational analysis of TatD DNase of Bacillus anthracis

Damini Singh  1 , Amit Rahi  1 , Romika Kumari  2 , Vatika Gupta  1 , Gunjan Gautam  3 , Somya Aggarwal  1 , Mohd Rehan  4 , Rakesh Bhatnagar  1
Affiliations

Computational and mutational analysis of TatD DNase of Bacillus anthracis

Damini Singh et al. J Cell Biochem. 2019 Jul.

Abstract

The role of TatD DNases as DNA repair enzymes or cell death (apoptotic) nucleases is well established in prokaryotes as well as eukaryotes. The current study aims to characterize the TatD nuclease from Bacillus anthracis (Ba TatD) and to explore its key histidine catalytic residues. Ba TatD was found to be a metal-dependent, nonspecific endonuclease which could efficiently cleave double-stranded DNA substrates. Moreover, Ba TatD nuclease was observed to be thermostable up to 55°C and act in a wide pH range indicating its industrial applicability. Diethyl pyrocarbonate-based histidine-selective alkylation of the Ba TatD resulted in a loss of its nuclease activity suggesting a crucial role of the histidine residues in its activity. The key residues of Ba TatD were predicted using sequence analysis and structure-based approaches, and then the predicted residues were further tested by mutational analysis. Upon mutational analysis, H128 and H153 have been found to be crucial for Ba TatD activity, though H153 seems to bear an important but a dispensable role for the Ba TatD nuclease. Ba TatD had a uniform expression in the cytosol of B. anthracis, which indicates a significant role of the protein in the pathogen's life cycle. This is the first study to identify and characterize the TatD DNase from B. anthracis and will be helpful in gaining more insights on the role of TatD proteins in Gram-positive bacteria where it remains unexplored.

Keywords: Bacillus anthracis; TatD nuclease; active site; histidine; thermostable.

PubMed Disclaimer

References

REFERENCES
    1. Li JY, Wang NA. Genome-wide mutagenesis of Xanthomonas axonopodis pv. citri reveals novel genetic determinants and regulation mechanisms of biofilm formation. PLoS One. 2011;6(7): e21804.
    1. Chang ZG, Jiang N, Zhang YY, et al. The TatD-like DNase of Plasmodium is a virulence factor and a potential malaria vaccine candidate. Nat Commun. 2016;7:11537.
    1. Chen L, Shen D, Sun N, Xu J, Wang W, Dou D. Phytophthora sojae TatD nuclease positively regulates sporulation and negatively regulates pathogenesis. MPMI. 2014a;27:1070-1080.
    1. Chen YC, Li CL, Hsiao YY, Duh Y, Yuan HS. Structure and function of TatD exonuclease in DNA repair. Nucleic Acids Res. 2014b;42:10776-10785.
    1. Gannavaram S, Debrabant A. Involvement of TatD nuclease during programmed cell death in the protozoan parasite Trypanosoma brucei. Mol Microbiol. 2012;83:926-935.

LinkOut - more resources