A family of genes with growth factor and adenosine deaminase similarity are preferentially expressed in the salivary glands of Glossina m. morsitans

Abstract

A cDNA library constructed from salivary glands of tsetse fly, Glossina morsitans morsitans (Diptera: Glossinidae), was differentially screened, and two related full-length cDNAs were molecularly characterized: tsetse salivary growth factor, TSGF-1 and TSGF-2. The cDNAs encode for open reading frames (ORFs) of 494 and 506 aa, respectively, and display an overall 45% amino acid identity and 61% similarity to one another. Both genes are preferentially expressed in the salivary glands of male and female adult flies. In addition to salivary glands, both transcripts can be detected from the gut tissue. Only transcripts specific for TSGF-2 are detected in ovary and testes tissues of adults as well as in puparia, while neither gene is expressed during the larval developmental stages. The N-terminal region of both putative proteins contains a hydrophobic sequence with secretory signal peptide characteristics, and analysis of proteins in saliva by Western blot indicates that both are secreted. Western blot analysis indicates that TSGF-1 is synthesized at significantly higher levels than TSGF-2. The deduced protein sequences of both cDNAs display extensive similarities to two other proteins: insect derived growth factor (IDGF) characterized from Sarcophaga peregrina with growth-factor activity, and atrial gland specific antigen (AGSA or MDSF) characterized from Apylasia californica. In addition to growth factor similarity, all four related proteins share the evolutionarily conserved amino acid residues associated with the enzymatic deamination of adenosine, which is shown here to be present in salivary gland extracts of tsetse. While both genes are present and expressed in G. m. morsitans and G. p. palpalis, only TSGF-1 is present in G. austeni. We present the molecular characteristics of the cDNAs, their genomic arrangement and their regulation of expression in different fly tissues and species. We discuss the potential role of these proteins in hemostatis and in African trypanosome transmission by different species of tsetse.