Molecular characterization of three gut genes from Glossina morsitans morsitans: cathepsin B, zinc-metalloprotease and zinc-carboxypeptidase

Abstract

Insect gut enzymes are involved in digestion of dietary proteins. Additionally, these enzymes have been implicated in the process of pathogen establishment in several insects including the tsetse fly (Diptera:Glossinidae), which is the vector for African trypanosomes. Both the male and female tsetse can transmit trypanosomes and are strict blood feeders during all stages of their development. Here, we describe the molecular characterization of three gut genes: cathepsin B (GmCatB), zinc-metalloprotease (GmZmp) and zinc-carboxypeptidase (GmZcp). The cDNA for GmCatB encodes a protein for 340 amino acids with a predicted molecular mass of 38.2 kDa, while the 854 bp GmZmp cDNA encodes a protein of 254 amino acids with a molecular mass of 29 kDa. The GmZcp cDNA is 1319 bp in length and has a 354 amino acids open reading frame for coding a 40 kDa protein. All three cDNAs have signal peptide sequences associated with their N-terminal domains and structure analysis indicates that GmCatB and GmZmp are expressed as zymogens with pro-domains proteolytically removed for activity. The activation domain associated with the carboxypeptidase sequences is lacking in GmZcp. While GmCatB transcription is constitutive, teneral flies express very low levels of transcripts for GmZmp and GmZcp prior to the first bloodmeal. Transcription of all genes is induced and remains high throughout the digestion cycle within a few hours following the first bloodmeal ingestion. Both GmCatB and GmZcp are parasite responsive, with the expression of both genes being higher in trypanosome infected flies.