Characterization of glutamine:: fructose-6-phosphate aminotransferase from the ixodid tick, Haemaphysalis longicornis, and its critical role in host blood feeding

Glutamine: fructose-6-phosphate aminotransferase (GFAT, EC2.6.1.16) is the first, and rate-limiting, enzyme in the hexosamine biosynthetic pathway, and is involved in the regulation of chitin biosynthesis and glycosylation of proteins. We report here the molecular characterization and potential functions of a novel GFAT (HIGFAT) from the ixodid tick Haemaphysalis longicornis. HIGFAT consists of 696 amino acids, possesses a class 11 glutamine aminotransferase domain and two sugar isomerase motifs, and has a close phylogenetic relationship to insect GFAT. HIGFAT was expressed at all stages of development and in multiple organs. The transcription levels in the cuticle and midgut were enhanced significantly by blood feeding during the first 3 days and decreased on the fifth day, while those in salivary glands maintained almost the same level during the first 3 days, and decreased to a rather low level at 5 days postinfestation. Endogenous HIGFAT was identified at all developmental stages and in multiple organs, such as epidermis, midgut epithelium, salivary gland, ovary, Malpigian's tubule and trachea. It was identified as a protein of 78.4 kDa using Western blot analysis. Following RNA interference of HIGFAT, engorgement by adult females was reduced significantly. One of the potential mechanisms for this effect may be that the inhibition of HIGFAT limits chitin biosynthesis, so disrupting cuticle growth and possibly peritrophic matrix formation during blood feeding. (c) 2006 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.