Ancestral sequence reconstruction has been widely used to test evolution-based hypotheses. The genome of the European tick vector, Ixodes ricinus, encodes for defensin peptides with diverse antimicrobial activities against distantly related pathogens. These pathogens include fungi, Gram-negative, and Gram-positive bacteria, i.e., a wide antimicrobial spectrum. Ticks do not transmit these pathogens, suggesting that these defensins may act against a wide range of microbes encountered by ticks during blood feeding or off-host periods. As demonstrated here, these I. ricinus defensins are also effective against the apicomplexan parasite Plasmodium falciparum. To study the general evolution of antimicrobial activity in tick defensins, the ancestral amino acid sequence of chelicerate defensins, which existed approximately 444 million years ago, was reconstructed using publicly available scorpion and tick defensin sequences (named Scorpions-Ticks Defensins Ancestor, STiDA). The activity of STiDA was tested against P. falciparum and the same Gram-negative and Gram-positive bacteria that were used for the I. ricinus defensins. While some extant tick defensins exhibit a wide antimicrobial spectrum, the ancestral defensin showed moderate activity against one of the tested microbes, P. falciparum. This study suggests that amino acid variability and defensin family expansion increased the antimicrobial spectrum of ancestral tick defensins.

Department of Bioresources Fraunhofer Institute for Molecular Biology and Applied Ecology Giessen Germany

Department of Bioresources Fraunhofer Institute for Molecular Biology and Applied EcologyGiessen Germany; Institute for Insect Biotechnology Justus Liebig University of GiessenGiessen Germany

Institute of Parasitology Biology Centre of the Czech Academy of Sciences České Budějovice Czech Republic

Institute of Parasitology Biology Centre of the Czech Academy of Sciences České Budějovice Czech Republic; Department of Virology Veterinary Research InstituteBrno Czech Republic

Institute of Parasitology Université Lille CNRS INSERM CHU Lille Institut Pasteur de Lille U1019 UMR 8204 Centre d'Infection et d'Immunité de Lille Lille France

Institute of Parasitology Université Lille CNRS INSERM CHU Lille Institut Pasteur de Lille U1019 UMR 8204 Centre d'Infection et d'Immunité de LilleLille France; Institute of Parasitology Biology Centre of the Czech Academy of Sciences České Budějovice Czech Republic; Faculty of Science University of South BohemiaČeské Budějovice Czech Republic

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