A Toll receptor from Manduca sexta is in response to Escherichia coli infection

Abstract

Genomic and cDNA sequences of a Toll receptor were cloned from the tobacco hornworm, Manduca sexta. M. sexta Toll (MsToll) gene contains six introns and seven exons. The full-length cDNA of MsToll is 3495 bp with an open reading frame of 2892 bp, which encodes a protein of 963 amino acids. MsToll is a typical single-pass transmembrane protein containing characteristic domain architecture of Toll and Toll-like receptors, including an extracellular domain composing of leucine-rich repeats (LRRs) and a cytoplasmic TIR domain. The deduced amino acid sequence of MsToll is most similar to Apis mellifera Toll (27% identity). Reverse-transcription polymerase chain reaction (RT-PCR) showed that MsToll was expressed in hemocytes, fat body, Malpighian tubule, midgut and epidermis. Real-time PCR analysis showed that MsToll mRNA in hemocytes was significantly induced by Escherichia coli, as well as by yeast (Saccharomyces cerevisiae) and Micrococcus lysodeikticus. However, MsToll transcript in fat body was not induced by these microorganisms. Immunohistochemistry assay showed that MsToll was detected on hemocytes. The TIR domain of MsToll also has high similarity to those of vertebrate TLR4 and zebra fish TLR (35-39% identity). Our results suggest that MsToll may play a role in innate signaling in response to E. coli infections.