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Purifying selection shaping the evolution of the Toll-like receptor 2 TIR domain in brown hares (Lepus europaeus) from Europe and the Middle East

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Abstract

Toll-like receptors (TLRs) are transmembrane proteins of the innate immune system, composed of the ectodomain involved in pathogen recognition and the intracellular Toll/interleukin-1 receptor (TIR) domain important for downstream signal transduction. Here, we analyze the genetic variability of TIR nucleotide and amino-acid sequences of the TLR2 gene in 243 brown hares from Europe and the Middle East and tested for the presence of selection signals and spatial structuring. TLR2 TIR domain sequences were PCR amplified and sequenced, while genotyping was performed by phasing. Genetic diversity indices were calculated in DnaSP and Arlequin, while presence of selection signals was tested using MEGA and the Datamonkey web server. The presence of spatial patterns in TIR sequence distribution was tested by spatial Principal Component Analysis (sPCA) in adegenet. A total of 13 haplotypes were revealed with haplotype diversity of 0.424, and nucleotide diversity (π) of 0.00138. Two spatial clusters were revealed: “Anatolia/Middle East” and “Europe”. In Anatolia the two most prevalent amino-acid variants, A and B (the latter being the most ancestral) were maintained at similar frequencies; but in Europe a shift in genotype frequencies was observed as well as a higher number of nonsynonymous substitutions giving rise to novel amino-acid protein variants originating from the evolutionarily younger protein variant. Molecular diversity (haplotype and nucleotide diversity) indices were significantly higher in the “Anatolia/Middle East” cluster. A signal of purifying selection was detected acting on the TIR sequences.

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Acknowledgements

We thank all local hunters, Prof. Philip U. Alkon, PhD (Israel), Orsolya Vincze, PhD (Romania) and Sim Broekhuizen, PhD (The Netherlands) for helping collecting the samples. Partial financial support for this work was provided by Wildlife Research—Franz Suchentrunk (Project No. 1/2018).

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Authors and Affiliations

  1. Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, Novi Sad, Serbia

    Milomir Stefanović, Mihajla Djan & Nevena Veličković

  2. Faculty of Science and Arts, University of Kırıkkale, Kırıkkale, Turkey

    Yasin Demirbaş

  3. Faculty of Forestry, Technical University, Zvolen, Slovakia

    Ladislav Paule

  4. Institute for Soil Sciences and Agricultural Chemistry, Hungarian Academy of Sciences, Budapest, Hungary

    Csongor István Gedeon

  5. Research Institute of Wildlife Ecology, University of Veterinary Medicine Vienna, Vienna, Austria

    Annika Posautz, Christoph Beiglböck, Anna Kübber-Heiss & Franz Suchentrunk

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  1. Milomir Stefanović
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  2. Mihajla Djan
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Stefanović, M., Djan, M., Veličković, N. et al. Purifying selection shaping the evolution of the Toll-like receptor 2 TIR domain in brown hares (Lepus europaeus) from Europe and the Middle East. Mol Biol Rep 47, 2975–2984 (2020). https://doi.org/10.1007/s11033-020-05382-x

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