A Lepidopteran-Specific Gene Family Encoding Valine-Rich Midgut Proteins
|BOMBYX-MORI; CHITIN SYNTHASE; CUTICULAR PROTEINS; EXPRESSION; INTESTINAL MUCIN; K+/H+ ANTIPORT; MANDUCA-SEXTA; Multidisciplinary Sciences; PERITROPHIC MATRIX; PLASMA-MEMBRANE; Science & Technology - Other Topics; TOBACCO HORNWORM
|PUBLIC LIBRARY SCIENCE
Many lepidopteran larvae are serious agricultural pests due to their feeding activity. Digestion of the plant diet occurs mainly in the midgut and is facilitated by the peritrophic matrix (PM), an extracellular sac-like structure, which lines the midgut epithelium and creates different digestive compartments. The PM is attracting increasing attention to control lepidopteran pests by interfering with this vital function. To identify novel PM components and thus potential targets for insecticides, we performed an immunoscreening with anti-PM antibodies using an expression library representing the larval midgut transcriptome of the tobacco hornworm, Manduca sexta. We identified three cDNAs encoding valine-rich midgut proteins of M. sexta (MsVmps), which appear to be loosely associated with the PM. They are members of a lepidopteran-specific family of nine VMP genes, which are exclusively expressed in larval stages in M. sexta. Most of the MsVMP transcripts are detected in the posterior midgut, with the highest levels observed for MsVMP1. To obtain further insight into Vmp function, we expressed MsVMP1 in insect cells and purified the recombinant protein. Lectin staining and glycosidase treatment indicated that MsVmp1 is highly O-glycosylated. In line with results from qPCR, immunoblots revealed that MsVmpl amounts are highest in feeding larvae, while MsVmpl is undetectable in starving and molting larvae. Finally using immunocytochemistry, we demonstrated that MsVmpl localizes to the cytosol of columnar cells, which secrete MsVmpl into the ectoperitrophic space in feeding larvae. In starving and molting larvae, MsVmpl is found in the gut lumen, suggesting that the PM has increased its permeability. The present study demonstrates that lepidopteran species including many agricultural pests have evolved a set of unique proteins that are not found in any other taxon and thus may reflect an important adaptation in the highly specialized lepidopteran digestive tract facing particular immune challenges.
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checked on Feb 22, 2024