Project description:Blood feeding animals face their host's defenses against tissue injury and blood loss while attempting to feed. One adaptation to surmount these barriers involves the evolution of a salivary potion that disarms their host's inflammatory and anti-hemostatic processes. The composition of the peptide moiety of this potion, or sialome (from the Greek sialo = saliva), can be deducted in part by proper interpretation of the blood feeder' sialotranscriptome. In this work we disclose the sialome of the blood feeding adult female Tabanus bromius. Following assembly of over 75 million Illumina reads (101 nt long) 16,683 contigs were obtained from which 4078 coding sequences were extracted. From these, 320 were assigned as coding for putative secreted proteins. These 320 contigs mapped 85% of the reads. The antigen-5 proteins family was studied in detail, indicating three Tabanus specific clades with and without disintegrin domains, as well as with and without leukotriene binding domains. Defensins were also detailed; a clade of salivary tabanid peptides was found lacking the propeptide domain ending in the KR dipeptide signaling furin cleavage. Novel protein families were also disclosed. Viral transcripts were identified closely matching the Kotonkan virus capsid proteins. Full length Mariner transposases were also identified. A total of 3043 coding sequences and their protein products were deposited in Genbank. Hyperlinked excel spreadsheets containing the coding sequences and their annotation are available at http://exon.niaid.nih.gov/transcriptome/T_bromius/Tbromius-web.xlsx (hyperlinked excel spreadsheet, 11 MB) and http://exon.niaid.nih.gov/transcriptome/T_bromius/Tbromius-SA.zip (Standalone excel with all local links, 360 MB). These sequences provide for a platform from which further proteomic studies may be designed to identify salivary proteins from T. bromius that are of pharmacological interest or used as immunological markers of host exposure.
Project description:Spiroplasma helicoides TABS-2T (DSM 22551) was isolated from the gut of a horsefly (Tabanus abactor) collected near Ardmore, Oklahoma, USA, in 1987. Here, we report the complete genome sequence of this bacterium to facilitate the investigation of its biology and the comparative genomics among Spiroplasma species.
Project description:Spiroplasma litorale TN-1(T) (DSM 21781) was isolated from the gut of a green-eyed horsefly (Tabanus nigrovittatus), collected at Ocracoke Island in North Carolina in 1983. Here, we report the complete genome sequence of this bacterium to facilitate the investigation of its biology.
Project description:From a large distance tabanid flies may find their host animal by means of its shape, size, motion, odour, radiance and degree of polarization of host-reflected light. After alighting on the host, tabanids may use their mechano-, thermo-, hygro- and chemoreceptors to sense the substrate characteristics. Female tabanids prefer to attack sunlit against shady dark host animals, or dark against bright hosts for a blood meal, the exact reasons for which are unknown. Since sunlit darker surfaces are warmer than shady ones or sunlit/shady brighter surfaces, the differences in surface temperatures of dark and bright as well as sunlit and shady hosts may partly explain their different attractiveness to tabanids. We tested this observed warmth preference in field experiments, where we compared the attractiveness to tabanids (Tabanus tergestinus) of a warm and a cold shiny black barrel imitating dark hosts with the same optical characteristics. Using imaging polarimetry, thermography and Schlieren imaging, we measured the optical and thermal characteristics of both barrels and their small-scale models. We recorded the number of landings on these targets and measured the time periods spent on them. Our study revealed that T. tergestinus tabanid flies prefer sunlit warm shiny black targets against sunlit or shady cold ones with the same optical characteristics. These results support our new hypothesis that a blood-seeking female tabanid prefers elevated temperatures, partly because her wing muscles are more rapid and her nervous system functions better (due to faster conduction velocities and synaptic transmission of signals) in a warmer microclimate, and thus, she can avoid the parasite-repelling reactions of host animals by a prompt take-off.
Project description:A novel family of RGD-containing molecules (Tablysin-15) has been molecularly characterised from the salivary gland of the haematophagous horsefly Tabanus yao. Tablysin-15 does not share primary sequence homology to any disintegrin discovered so far, and displays an RGD motif in the N-terminus of the molecule. It is also distinct from disintegrins from Viperidae since its mature form is not released from a metalloproteinase precursor. Tablysin-15 exhibits high affinity binding for platelet ?IIb?3 and endothelial cell ?V?3 integrins, but not for ?5?1 or ?2?1. Accordingly, it blocks endothelial cell adhesion to vitronectin (IC50 ~1 nM) and marginally to fibronectin (IC50 ~1 ?M), but not to collagen. It also inhibits fibroblast growth factor (FGF)-induced endothelial cell proliferation, and attenuates tube formation in vitro. In platelets, Tablysin-15 inhibits aggregation induced by collagen, ADP and convulxin, and prevents static platelet adhesion to immobilised fibrinogen. In addition, solid-phase assays and flow cytometry demonstrates that ?IIb?3 binds to Tablysin-15. Moreover, immobilised Tablysin-15 supports platelet adhesion by a mechanism which was blocked by anti-integrin ?IIb?3 monoclonal antibody (e.g. abciximab) or by EDTA. Furthermore, Tablysin-15 dose-dependently attenuates thrombus formation to collagen under flow. Consistent with these findings, Tablysin-15 displays antithrombotic properties in vivo suggesting that it is a useful tool to block ?IIb?3, or as a prototype to develop antithrombotics. The RGD motif in the unique sequence of Tablysin-15 represents a novel template for studying the structure-function relationship of the disintegrin family of inhibitors.