Evolutionary Divergence of CXCR1 (Interleukin-8 Receptor A) Gene of Indian Water Buffalo (Bubalus bubalis) in Light of Molecular Evolution

  • Shiveeli Rajput
  • Jaspreet Singh AroraEmail author
  • Chandra Sekhar Mukhopadhyay
  • Jasdeep Kaur Dhanoa
  • Ramneek Verma
  • R. S. Sethi
  • Satparkash Singh
  • B. V. Sunil Kumar
  • Sikh Tejinder Singh
Research Article


CXCR1 (Interleukin-8 Receptor A) is one of the two high-affinity receptors (IL-8RA and IL-8RB), present on the surface of neutrophils that binds to the IL-8 ligand, which results in the activation of immune response and allows the migration of immune cells to the site of inflammation. CXCR1 is strongly associated with inflammatory responses against most of the Gram-negative bacteria. The objective of the present work was to amplify, sequence and in silico characterize CXCR1 coding region of buffalo. The amplified target gene of buffalo CXCR1 was custom-sequenced, and the annotated sequence of 1130 bp having coding region of 1083 bp was submitted to DDBJ with accession number LC384988. Pairwise sequence alignment of buffalo CXCR1 and the reference cattle CXCR1 showed 95% similarity. Sequence analysis and integrative analysis of CXCR1 in 36 divergent species revealed that 7 codon positions have undergone positive selection pressure, indicating that the gene has undergone evolutionary changes over the period of time. Evolutionary analysis of the CXCR1 (MEGA 6 and Datamonkey online server) revealed that ruminant, avian, fish CXCR1 formed different clusters within the phylogenetic tree, showing evolutionary divergence.


CXC-Chemokine receptors Interleukin-8 Receptor A Sequence analysis Characterization 



Financial assistance for conducting the research work provided by UGC and RKVY is highly acknowledged.

Compliance with Ethical Standards

Conflict of interest

All authors declare that they have no conflict of interest to publish this manuscript.


  1. 1.
    Beecher C, Daly M, Childs S, Berry DP, Magee DA, McCarthy TV, Giblin L (2010) Polymorphisms in bovine immune genes and their associations with somatic cell count and milk production in dairy cattle. BMC Genet 11(1):99–102CrossRefGoogle Scholar
  2. 2.
    Abera M, Demie B, Aragaw K, Regassa F, Regassa A (2010) Isolation and identification of Staphylococcus aureus from bovine mastitic milk and their drug resistance patterns in Adama Town, Ethiopia. J Vet Med Anim Health 2(3):29–34Google Scholar
  3. 3.
    Chuntharapai A, Lee J, Hebert CA, Kim KJ (1994) Monoclonal antibodies detect different distribution patterns of IL-8 receptor A and IL-8 receptor B on human peripheral blood leukocytes. J Immunol 153(12):5682–5688PubMedGoogle Scholar
  4. 4.
    Burvenich C, Paape MJ, Hill AW, Guidry AJ, Miller RH, Heyneman R, Brand A (1994) Role of the neutrophil leucocyte in the local and systemic reactions during experimentally induced E. coli mastitis in cows immediately after calving. Vet Q 16(1):45–50CrossRefGoogle Scholar
  5. 5.
    Oviedo-Boyso J, Valdez-Alarcón JJ, Cajero-Juárez M, Ochoa-Zarzosa A, López-Meza JE, Bravo-Patino A, Baizabal-Aguirre VM (2007) Innate immune response of bovine mammary gland to pathogenic bacteria responsible for mastitis. J Infect 54(4):399–409CrossRefGoogle Scholar
  6. 6.
    Rainard P, Riollet C (2006) Innate immunity of the bovine mammary gland. Vet Res 37(3):369–400CrossRefGoogle Scholar
  7. 7.
    Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215(3):403–410CrossRefGoogle Scholar
  8. 8.
    Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol 35(6):1547–1549CrossRefGoogle Scholar
  9. 9.
    Hasegawa M, Kishino H, Yano T (1985) Dating of the human-ape splitting by a molecular clock of mitochondrial DNA. J Mol Evol 22(2):160–174. CrossRefPubMedGoogle Scholar
  10. 10.
    Tamura K, Kumar S (2002) Evolutionary distance estimation under heterogeneous substitution pattern among lineages. Mol Biol Evol 19(10):1727–1736CrossRefGoogle Scholar
  11. 11.
    Tamura K, Nei M, Kumar S (2004) Prospects for inferring very large phylogenies by using the eighbour-joining method. Proc Natl Acad Sci USA 101(30):11030–11035CrossRefGoogle Scholar
  12. 12.
    Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30(12):2725–2729CrossRefGoogle Scholar
  13. 13.
    Pond SLK, Frost SD (2005) Datamonkey: rapid detection of selective pressure on individual sites of codon alignments. Bioinformatics 21(10):2531–2533CrossRefGoogle Scholar
  14. 14.
    Chen C, Li Z, Zhou Z, Yin Z, Chan SM, Yu XQ, He J (2009) Cloning, characterization and expression analysis of a CXCR1-like gene from mandarin fish Siniperca chuatsi. Fish Physiol Biochem 35(3):489–499CrossRefGoogle Scholar
  15. 15.
    Dige MS, Ahlawat SPS, Bhushan B, Kumar A, Kumar P, Dhama K, Bhatt P (2015) Molecular characterization of Exon-2 of CXCR2 Gene in Vrindavani Cattle. Vet Pract 16(2):291–293Google Scholar
  16. 16.
    Hedges SB (1992) The number of replications needed for accurate estimation of bootstrap P value in Phylogenetic studies. Proc R Soc 276:4261–4270Google Scholar
  17. 17.
    Widdison S, Siddiqui N, Easton V, Lawrence F, Ashley G, Werling D, Watson M, Coffey TJ (2010) The bovine chemokine receptors and their mRNA abundance in mononuclear phagocytes. BMC Genom 11:439CrossRefGoogle Scholar
  18. 18.
    Liu Y, Yang S, Lin AA, Cavalli-Sforza LL, Su B (2005) Molecular evolution of CXCR1, a G protein-coupled receptor involved in signal transduction of neutrophils. J Mol Evol 61(5):691–696CrossRefGoogle Scholar
  19. 19.
    Dhaliwal KK, Arora JS, Mukhopadhyay CS, Dubey PP (2015) In silico characterization of functional divergence of two cathelicidin variants in Indian sheep. Evolut Bioinform Online 11:189Google Scholar
  20. 20.
    Xu T, Zhu Z, Sun Y, Ren L, Wang R (2014) Characterization and expression of the CXCR1 and CXCR4 in miiuy croaker and evolutionary analysis shows the strong positive selection pressures imposed in mammal CXCR1. Dev Comp Immunol 44(1):133–144CrossRefGoogle Scholar

Copyright information

© The National Academy of Sciences, India 2018

Authors and Affiliations

  • Shiveeli Rajput
    • 1
  • Jaspreet Singh Arora
    • 1
    Email author
  • Chandra Sekhar Mukhopadhyay
    • 1
  • Jasdeep Kaur Dhanoa
    • 1
  • Ramneek Verma
    • 1
  • R. S. Sethi
    • 1
  • Satparkash Singh
    • 1
  • B. V. Sunil Kumar
    • 1
  • Sikh Tejinder Singh
    • 2
  1. 1.School of Animal BiotechnologyGuru Angad Dev Veterinary and Animal Sciences University (GADVASU)LudhianaIndia
  2. 2.Department of Animal Breeding and GeneticsGuru Angad Dev Veterinary and Animal Sciences UniversityLudhianaIndia

Personalised recommendations