Expression cloning and analysis of a putative Brugia malayi POU-homeodomain transcription factor

Abstract

Neglected tropical diseases (NTDs) are a series of diseases that afflict individuals living predominantly in developing countries. One of the most impactful NTDs is lymphatic filariasis (LF), a disease caused by filarial parasitic nematodes. Current efforts to combat these diseases have been met with considerable success, but the concern of possible drug resistance underscores a need for the development of novel anthelmintic therapeutics that cause minimal harm to the human host. Expression of the parasite’s genome undergoes dramatic changes when the parasite is transmitted from the mosquito vector into the human host. Despite the critical need to understand these gene expression changes, little is known about promoters and transcription factors in filarial parasites. The aim of this study is to identify transcription factors in our model organism, B. malayi, that could serve as potential drug targets to help combat filariasis. Through a comparative analysis between whole genome datasets of B. malayi and C. elegans, we were able to identify the putative transcription factor Bma-UNC-86 as the best candidate for this initial project. Bioinformatics analyses revealed high homology between the binding sequences of UNC-86 in the promoter region of its target gene, mec-3, in both C. elegans and B. malayi. The putative transcription factor Bma-UNC-86 was cloned and expressed in E. coli. The purified UNC-86 protein will be used in future studies to investigate gene regulation at the protein level by analyzing the interaction of the transcription factor with the promoter region of its target gene in B. malayi. Application of this approach will further expand the data available to investigate the biology of parasitic helminths and lead to the development of novel therapeutics