Blood feeding tightly regulates the reproductive cycles in ticks. Vitellogenesis and nutritional signaling are a key event in the tick reproductive cycle. Here we report the identification of a Haemaphysalis longicornis GATA factor, (HlGATA), which is synthesized after a blood meal and acts as a transcriptional activator of vitellogenin (Vg). HlGATA shares structural similarity with other GATA factors of invertebrates and vertebrates. Tick GATA mRNA accumulated in the fat body and midgut prior to blood feeding. However, translation of GATA was activated by blood feeding because the GATA protein increased dramatically in engorged females. RNA interference (RNAi)-mediated knock down of GATA transcript resulted in a significant inhibition of Vg expression and effectively disrupts egg development after blood meal in engorged tick. In addition, HlGATA translation was inhibited by RNAi-mediated knock down of S6 kinase. These experiments have revealed that the GATA factor, which is the specific transcriptional activator of Vg gene, represents important molecule for tick reproduction.

Antimicrobial peptides represent an essential alternative first line of defense. Naturally occurring molecules associated with the innate immune system in disease-bearing vectors such as mosquito, tick could be the target for searching more potent and effective agents to combat against the pathogens resistant to conventionally used antibiotics. Recently, we explored expression of a defensinlike peptide, longicin from the hard tick Haemaphysalis longicornis. Longicin and one of its synthetic partial analog (P4) displayed antimicrobial/fungicidal/parasiticidal activity and, therefore, proposed to be a chemotherapeutic compounds against tick-borne disease organisms. Structural characterization of antimicrobial peptides is very important to understand the peptide activity. In addition, harmful side effects such as lysis of red blood cells or cytotoxicity towards mammalian host cells commonly associated with antimicrobial peptides as potential therapeutic agent should also be elucidated. In this study, we analyzed some structural features using bioinformatics tool, CD Spectroscopy, and also determined cytolytic activity of P4 peptide. According to the chemicophysical characteristics, the P4 is suggested to be a cationic peptide with hydrophobic and amphipathic character. The predicted secondary structure indicated the existence of β-sheet which was also observed in modelled tertiary structure. CD spectroscopy results also revealed the existence of a β-sheet and changes of helical content in the presence of membrane-mimic condition. These structural observations on P4 suggest that the antimicrobial activity could be due to the well developed β-sheet. In addition, sequence homology search showed that antimicrobial molecule identified in other ticks and in organisms have the P4 analogous domain at their C-terminal, which indicates P4 as a conserved antimicrobial domain. The peptide P4 also showed less cytolytic activity against various cell lines or erythrocytes of various species. The data presented here strongly suggests that the peptide P4 could be developed as future therapeutic agent against tick-borne microorganisms.