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Molecular characterization of coding sequences and analysis of Toll-like receptor 3 mRNA expression in water buffalo (Bubalus bubalis) and nilgai (Boselaphus tragocamelus)

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Abstract

Toll-like receptor 3 (TLR3), an antiviral innate immunity receptor recognizes double-stranded RNA, preferably of viral origin and induces type I interferon production, which causes maturation of phagocytes and subsequent release of chemical mediators from phagocytes against some viral infections. The present study has characterized TLR3 complementary DNA (cDNA) in buffalo (Bubalus bubalis) and nilgai (Boselaphus tragocamelus). TLR3 coding sequences of both buffalo and nilgai were amplified from cultured dendritic cell cDNA and cloned in pGEMT-easy vector for characterization by restriction endonucleases and nucleotide sequencing. Sequence analysis reveals that 2,715-bp-long TLR3 open reading frame encoding 904 amino acids in buffalo as well as nilgai is similar to that of cattle. Buffalo TLR3 has 98.6 and 97.9% identity at nucleotide level with nilgai and cattle, respectively. Likewise, buffalo TLR3 amino acids share 96.7% identity with cattle and 97.8% with nilgai. Non-synonymous substitutions exceeding synonymous substitutions indicate evolution of this receptor through positive selection among these three ruminant species. Buffalo and nilgai appear to have diverged from a common ancestor in phylogenetic analysis. Predicted protein structures of buffalo and nilgai TLR3 from deduced amino acid sequences indicate that the buffalo and nilgai TLR3 ectodomain may be more efficient in ligand binding than that of cattle. Furthermore, TLR3 messenger RNA expression in tissues as quantified by real-time PCR was found higher in nilgai than buffalo.

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Acknowledgement

The authors are thankful to the Director of the Indian Veterinary Research Institute, Izatnagar for providing necessary facilities to carry out the work. A. D. is thankful to ICAR for providing the Junior Research Fellowship.

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Authors and Affiliations

  1. Centre for Wildlife, Indian Veterinary Research Institute, Izatnagar, Bareilly, 243122, India

    Animesh Dhara, Mohini Saini, Dhanjit K. Das & Devendra Swarup

  2. Division of Biochemistry, Indian Veterinary Research Institute, Izatnagar, Bareilly, 243122, India

    Bhaskar Sharma

  3. Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, Bareilly, 243122, India

    Satish Kumar & Praveen K. Gupta

Authors
  1. Animesh Dhara
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  2. Mohini Saini
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  3. Dhanjit K. Das
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  4. Devendra Swarup
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  5. Bhaskar Sharma
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  6. Satish Kumar
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  7. Praveen K. Gupta
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Correspondence to Mohini Saini.

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Photographs

(A, B) Buffalo Dendritic cells; C Immature Dendritic cell; D Mature Dendritic cell (E, F) Nilgai Dendritic cells. Dendritic cells are marked by arrow (DOC 10718 kb)

Table 1

Changes in the nucleotide positions in the TLR3 coding sequences (DOC 184 kb)

Table 2

Changes in the amino acid positions in different domain of TLR3 protein sequences (DOC 114 kb)

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Dhara, A., Saini, M., Das, D.K. et al. Molecular characterization of coding sequences and analysis of Toll-like receptor 3 mRNA expression in water buffalo (Bubalus bubalis) and nilgai (Boselaphus tragocamelus). Immunogenetics 59, 69–76 (2007). https://doi.org/10.1007/s00251-006-0174-6

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  • DOI: https://doi.org/10.1007/s00251-006-0174-6

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