Novel SNPs of the ABCG2 Gene and Their Associations with Milk Production Traits in Iranian Holstein Bulls

Authors
S. A. Mousavizadeh 1
A. Salehi 1
M. Aminafshar 2
M. Sayyadnejad 3
M. H. Nazemshirazi 4
1 Department of Animal and Poultry Science, Abureyhan Campus, University of Tehran, Pakdasht, Islamic Republic of Iran.
2 Department of Animal Science, Faculty of Agriculture and Natural Resources, Science and Research Branch, Islamic Azad University, Tehran, Islamic Republic of Iran.
3 Animal Breeding Center of Iran, Karaj, Islamic Republic of Iran.
4 Department of Molecular, Iran Central Veterinary Laboratory, Tehran, Islamic Republic of Iran.
Abstract
ABCG2 (ATP binding cassette subfamily G member 2) gene, located on chromosome 6 encodes the ABCG2 protein that transports various xenobiotics, cytostatic drugs across the plasma membrane as well as cholesterol into milk. A single nucleotide change (A/C) in base 86 of exon 14 is capable of encoding a substitution of tyrosine with serine in the ABCG2 gene and increase milk yield while decreasing milk fat and protein concentrations. The major aim followed in this research was to study Single Nucleotide Polymorphisms (SNPs) of ABCG2 gene and their association with milk production traits in Iranian Holstein bulls. Genomic DNA of 105 ified bulls was extracted from semen samples using highly Pure PCR template preparation kit. Primers were designed through Oligo software (Version 5.0) and utilized in PCR. Then the PCR fragments were sequenced. The A/C substitution in base number 86 of exon 14 was observed with 2% frequency which affected protein percentage (P< 0.05). Some SNPs were detected for the first time in intron 13, exon and intron 14 in comparison with sequences in the NCBI database. A deletion mutation in base number 20 (T/-) and a missense mutation in base number 67 (A/G) of exon 14 that cause the substitution of serine with glycine were discovered which were significantly associated with protein yield and fat percentage, respectively (P< 0.05). Furthermore, significant association was observed between fat percentage and mutations in base numbers 4,133 (T/C) and 4,137 (T/G) of intron 13 (P< 0.05). Substitutions in base numbers 2 (T/C) and 55 (G/C) of intron 14 resulted in a significant effect on fat yield and fat percentage (P< 0.05).

Keywords

1. Cohen, M., Seroussi, E. and Band, M. 2004. SPP1 is a candidate gene for the QTL affecting milk protein concentration on BTA6 based on population-wide linkage disequilibrium, differential expression, and targeted inhibition. 29th Conference of the International Society for Animal Genetics, Tokyo, Japan.
2. Cohen-Zinder, M., Seroussi, E., Larkin, D. M., Loor, J. J., Wind, A. E., Lee, J. H., Drackley, J. K., Band, M. R., Hernandez, A. G., Shani, M., Lewin, H. A., Weller, J. I., and Ron, M. 2005. Identification of a missense mutation in the bovine ABCG2 gene with a major effect on the QTL on chromosome 6 affecting milk yield and composition in Holstein cattle. Genome Res. 15: 936–44.
3. Farke, C., Meyer, H. H., Bruckmaier, R. M. and Albrecht, C. 2008. Differential expression of ABC transporters and their regulatory genes during lactation and dry period in bovine mammary tissue. J Dairy Res. 75: 406–414.
4. Jonker, J. W., Merino, G., Musters, S., Van Herwaarden, A. E., Bolscher, E., Wagenaar, E., Mesman, E., Dale, T. C., and Schinkel, A. H. 2005. The breast cancer resistance protein BCRP (ABCG2) concentrates drugs and carcinogenic xenotoxins into milk. Nat. Med. 11: 127–129.
5. Khatib, H., Zaitoun, I., Wiebelhaus-Finger, J., Chang, Y. M. and Rosa, G. J. M. 2007. The Association of Bovine PPARGC1A and OPN Genes with Milk Composition in Two Independent Holstein Cattle Populations. J. Dairy Sci. 90: 2966–2970.
6. Komisarek, J., Dorynek, Z. 2009. Effect of ABCG2, PPARGC1A, OLR1 and SCD1 gene polymorphism on estimated breeding values for functional and production traits in Polish Holstein-Friesian bulls. J. Appl Genet. 50(2): 125–132.
7. Leonard, S., Khatib, H., Schutzkus, V., Chang, Y. M. and Maltecca, C. 2005. Effects of the Osteopontin gene variants on milk production traits in dairy cattle. J. Dairy Sci. 88:4083–4086.
8. Leslie, E. M., Deeley, R. G. and Cole, S. P. 2005. Multidrug resistance proteins: role of P-glycoprotein, MRP1, MRP2, and BCRP (ABCG2) in tissue defense. Toxicology and Applied Pharmacology. 204: 216–237.
9. Litman, T., Brangi, M., Hudson, E., Fetch, P., Abati, A., Ross, D. D., Miyake, K., Resau, J. H. and Bates, S. E. 2000. The multidrugresistant phenotype associated with overexpression of the new ABC half-transporter, MXR (ABCG2). J. Cell Sci. 113:2011–2021.
10. Olsen, H. G., Lien, S., Gautier, M., Nilsen, H., Roseth, A., Berg, P. R., Sundsaasen, K. K., Svendsen, M. and Meuwissen, T. H. E. 2005. Mapping of a milk production quantitative trait locus to a 420-kb region on bovine chromosome 6. Genetics. 169: 275–83.
11. Olsen, H. G., Nilsen, H., Hayes, B., Berg, P. R., Svendsen, M., Lien, S. and Meuwissen, T. 2007. Genetic support for a quantitative trait nucleotide in the ABCG2 gene affecting the milk composition of dairy cattle. BMC Genetics. 8, 32.
12. Ron, M., Cohen-Zinder, M., Peter, C., Weller, J. I. and Erhardt, G. 2006). Short Communication: A Polymorphism in ABCG2 in Bos indicus and Bos Taurus Cattle Breeds. J. Dairy Sci. 89:4921–4923.
13. Sarkadi, B., OzvegyLaczka, C., Nemet, K., Varadi, A. 2004. Minireview ABCG2 – a transporter for all seasons. FEBS Letters. 567 (2004) 116–120.
14. Schnabel, R. D., Kim, J. J., Ashwell, M. S., Sonstegard, T. S., Van Tassell, C. P., Connor, E.E., and Taylor, J. F. 2005. Fine-mapping milk production quantitative trait loci on BTA6: Analysis of the bovine Osteopontin gene. Proc. Natl. Acad. Sci. USA. 102:6896–6901.
15. Van Herwaarden, A. E.,Wagenaar, E., Merino, G., Jonker, J. W., Rosing, H., Beijnen, J. H. and Schinkel, A. H. 2007. Multidrug transporter ABCG2/breast cancer resistance protein secretes riboflavin (vitamin B2) into milk. Mol Cell Biol. 27: 1247–1253.
16. Weikard, R., Kuhn, C., Goldammer, T., Freyer, G. and Schwerin, M. 2005. The bovine PPARGC1A gene: molecular characterization and association of an SNP with variation of milk fat synthesis. Physiological Genomics. 21: 1–3.
17. Zhou, S., Schuetz, J. D., Bunting, K. D., Colapietro, A. M., Sampath, J., Morris, J. J., Lagutina, I., Grosveld, G. C., Osawa, M., Nakauchi, H. and Sorrentino, B. P. 2001. The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype. Nat Med. 7:1028–1034.
Journal of Agricultural Science and Technology
Volume 15, Issue 6
November and December 2013
Pages 1145-1151