Genetic analysis of glycosylation sites, TM 8 and consensus transport motif of buffalo SLC11A1 gene

The present study aimed to explore polymorphisms at exon X-XI, encompassing two potential N-linked Glycosylation sites (GLY, GLY), complete transmembrane domain 8 (TM8) and one conserved ‘Consensus Transport Motif ’(CTM) of buffalo (Bubalus bubalis) SLC11A1 (formerly NRAMP1) gene by PCR-RFLP and automated DNA sequencing. PCR-RFLP genotyping of animals (n=150) from three Indian buffalo breeds viz., Murrah, Surti, and Mehsana by Alu I, Taq I, Hae III, Rsa I, Pst I and Mva I reveals monomorphic pattern. Sequence comparisons demonstrate no change at nucleotide level among the buffalo breeds. However, results of this study show three nucleotide changes in buffaloes as compared to that of B taurus (NM_174652): one transition (C-»T at amino acid position 349) and two transversion (C-»A at amino acid position 321 and C-»G at amino acid position 356). Transversion C-»Aand C-»Q resulting in deduced amino acid exchange Asp₃₂₁-Glu₃₂₁ and Pro₃₅₆-Ala₃₅₆, respectively has been identified. The non-synonymous single nucleotide variation (nsSNV) (C-»A at amino acid position 321), observed in all the three buffalo breeds under study, leads to the loss of a potential N-glycosylation motif of SLC11A1 gene and therefore may alter the property of the protein. A comprehensive study of this N-glycosylation motif of SLC11A1 gene in different species followed by structural analysis of the protein is required to better understand the functional impact of nsSNV.