Cloning, characterization, 3D modeling and dsRNA studies of Aedes aegypti juvenile hormone acid methyl transferase

Juvenile hormone acid methyl transferase (JHAMT), the ultimate enzyme in the JH III biosynthetic pathway in the mosquito Aedes aegypti, is an important key enzyme that converts JH acid (JHA) into JHIII. Disrupting JH bio synthetic pathway can be used to control insects that vectors many arboviruses such as, dengue, chikungunya and yellow fever. Thus, the JHAMT gene of Ae.aegypti (jmtA) was sequenced, the sequence deposited in the GenBank (accession #DQ409061), the enzyme expressed and characterized using different substrates and the K_m and V_max of the different substrates were calculated using Lineweaver Burk reciprocal plots. To study the relationship between binding and substrate specificity a 3D model was built and docking of the substrate to the enzyme was studied to investigate substrate binding on the enzyme surface predicated from our 3D model. RNA mediated interference (RNAi) studies using jmtA dsRNA inhibited egg development in female Ae. aegypti and delayed adult emergence and larval development for up to 3 weeks past normal pupation time. Following pupation, the newly emerged adults died without taking a blood meal. These results and related findings will be discussed.