Abstract
The lymphatic filarial parasite, Brugia malayi contains Wolbachia endobacteria that are essential for development, viability and fertility of the parasite. Therefore, wolbachial proteins have been currently seen as the potential antifilarial drug targets. NAD+-dependent DNA ligase is characterized as a promising drug target in several organisms due to its crucial, indispensable role in DNA replication, recombination and DNA repair. We report here the cloning, expression and purification of NAD+-dependent DNA ligase of Wolbachia endosymbiont of B. malayi (wBm-LigA) for its molecular characterization. wBm-LigA has all the domains that are present in nearly all the eubacterial NAD+-dependent DNA ligases such as N-terminal adenylation domain, OB fold, helix-hairpin-helix (HhH) and BRCT domain except zinc-binding tetracysteine domain. The purified recombinant protein (683-amino acid) was found to be biochemically active and was present in its native form as revealed by the circular dichroism and fluorescence spectra. The purified recombinant enzyme was able to catalyze intramolecular strand joining on a nicked DNA as well as intermolecular joining of the cohesive ends of BstEII restricted lamda DNA in an in vitro assay. The enzyme was localized in the various life-stages of B. malayi parasites by immunoblotting and high enzyme expression was observed in Wolbachia within B. malayi microfilariae and female adult parasites along the hypodermal chords and in the gravid portion as evident by the confocal microscopy. Ours is the first report on this enzyme of Wolbachia and these findings would assist in validating the antifilarial drug target potential of wBm-LigA in future studies.
Highlights
More than 1.3 billion people in 81 countries worldwide are threatened by lymphatic filariasis (LF)
A few proteins of Wolbachia have been identified and their functions are being investigated such as Wolbachia surface protein (WSP), heat shock protein 60 (HSP60), independent phosphoglycerate mutase and the enzymes involved in Heme biosynthetic pathway
DNA ligase of Wolbachia plays an indispensable role in the intracellular bacterial division inside the B. malayi parasite
Summary
More than 1.3 billion people in 81 countries worldwide are threatened by lymphatic filariasis (LF). Characterization of novel proteins of Wolbachia is a prerequisite to identify the factors responsible for pathogenesis of the disease, understanding the host- parasite interactions and developing new protein/enzyme targets for antifilarial chemotherapy. DNA ligases are indispensable for many fundamental processes in DNA metabolism including the linkage of Okazaki fragments during replication, recombination processes and repair pathways requiring resynthesis of DNA [4,5]. Their crucial function is emphasized by the fact that the eukaryotic cells contain several isoenzymes and that the viruses encode their own ligases [6]. To the best of our knowledge, this is the first ever report on the characterization of any nuclear metabolic enzyme from an obligate mutualist alphaproteobacterium, Wolbachia of filarial parasite
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
