During the blood feeding, sand fly females inject saliva containing immunomodulatory and anti-haemostatic molecules into their vertebrate hosts. The saliva composition is species-specific, likely due to an adaptation to particular haemostatic pathways of their preferred host. Research on sand fly saliva is limited to the representatives of two best-studied genera, Phlebotomus and Lutzomyia. Although the members of the genus Sergentomyia are highly abundant in many areas in the Old World, their role in human disease transmission remains uncertain. Most Sergentomyia spp. preferentially attack various species of reptiles, but feeding on warm-blooded vertebrates, including humans and domestic animals, has been repeatedly described, especially for Sergentomyia schwetzi, of which salivary gland transcriptome and proteome is analyzed in the current study. Illumina RNA sequencing and de novo assembly of the reads and their annotation revealed 17,293 sequences homologous to other arthropods' proteins. In the sialome, all proteins typical for sand fly saliva were identified-antigen 5-related, lufaxin, yellow-related, PpSP15-like, D7-related, ParSP25-like, and silk proteins, as well as less frequent salivary proteins included 71kDa-like, ParSP80-like, SP16-like, and ParSP17-like proteins. Salivary enzymes include apyrase, hyaluronidase, endonuclease, amylase, lipase A2, adenosine deaminase, pyrophosphatase, 5'nucleotidase, and ribonuclease. Proteomics analysis of salivary glands identified 631 proteins, 81 of which are likely secreted into the saliva. We also compared two S. schwetzi lineages derived from the same origin. These lineages were adapted for over 40 generations for blood feeding either on mice (S-M) or geckos (S-G), two vertebrate hosts with different haemostatic mechanisms. Altogether, 20 and 40 annotated salivary transcripts were up-regulated in the S-M and S-G lineage, respectively. Proteomic comparison revealed ten salivary proteins more abundant in the lineage S-M, whereas 66 salivary proteins were enriched in the lineage S-G. No difference between lineages was found for apyrase activity; contrarily the hyaluronidase activity was significantly higher in the lineage feeding on mice.

• MeSH
• apyrasa analýza   genetika   metabolismus   MeSH
• fylogeneze MeSH
• hmyzí proteiny analýza   genetika   metabolismus   MeSH
• hyaluronoglukosaminidasa analýza   genetika   metabolismus   MeSH
• ještěři MeSH
• myši MeSH
• Psychodidae genetika   metabolismus   MeSH
• receptory pachové analýza   genetika   metabolismus   MeSH
• slinné žlázy metabolismus   MeSH
• transkriptom * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Ecto-5'-nucleotidases are glycosyl phosphatidylinositol (GPI)-linked membrane-bound glycoproteins that convert extracellular AMP to adenosine. They play important roles in the inflammatory response where they modulate levels of pro-inflammatory extracellular ATP and anti-inflammatory extracellular adenosine. They are found in the saliva of blood feeding insects and also have a role in male reproduction. Drosophila possesses five genes with eight alternative transcripts encoding proteins with sequence homology to mammalian ecto-5'-nucleotidases. Here we show that two of them - NT5E-1 (CG4827) and NT5E-2 (CG30104) - are GPI-linked proteins with ecto-5'-nucleotidase activity but that they can also be released from the GPI anchor and exhibit secreted 5'-nucleotidase activity in growth media. The third locus in the cluster, CG30103, most likely also encodes a GPI-anchored membrane-bound protein but without 5'-nucleotidase activity, possibly due to the numerous substitutions in the amino acid sequence. Together with NT5E-2, CG30103 is also expressed in the testis offering an interesting model to investigate ecto-5'-nucleotidase enzymatic and extra-enzymatic function in male reproduction. CG42249 locus encoding two alternative transcripts is sequentially similar to family of apyrases related to 5'-nucleotidases and we show here that together with CG5276 belonging to another family of calcium-activated nucleotidases function as apyrases converting extracellular ATP to ADP and AMP. The last locus, CG11883, encodes most likely a cytoplasmic/mitochondrial protein.

• MeSH
• 5'-nukleotidasa genetika   metabolismus   MeSH
• apyrasa genetika   metabolismus   MeSH
• buněčné linie MeSH
• Drosophila melanogaster enzymologie   genetika   MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• multigenová rodina MeSH
• proteiny Drosophily genetika   metabolismus   MeSH
• sekvence aminokyselin MeSH
• vývojová regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH