Skip to main content

Advertisement

SpringerLink
Log in

Cloning, Sequence and Functional Analysis of Goat ATP-binding Cassette Transporter G2 (ABCG2)

  • Research
  • Published:
Molecular Biotechnology Aims and scope Submit manuscript

Abstract

ATP-binding cassette transporter G2 (ABCG2) gene encodes a protein that has a wide variety of substrates and is responsible for the active secretion of clinically and toxicologically important molecules into milk. Although known in many species, this marks the first time this gene product has been reported in goats. In this study, we cloned and sequenced goat ABCG2 gene complete coding sequence and predicted its putative translated protein structure with implicative functional domains. One six-transmembrane span on C-terminal region and at least one coiled-coil domain on N-terminal were predicted and compared primarily with those of other closely related species. In addition, three conserved cysteines (in positions 595, 606, and 611) were determined toward the C-terminal of goat’s ABCG2. Two known functional motifs were identified in goat’s protein through comparative studies with other species. The goat ABCG2 relative expression profile revealed that the gene expression was a function of lactation stage and parallel to goat lactation curve.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Sarkadi, B., Ozvegy-Laczka, C., Nemet, K., & Varadi, A. (2004). ABCG2-a transporter for all seasons. FEBS Letters, 567, 116-120.

    Article  PubMed  CAS  Google Scholar 

  2. Allikmets, R., Wasserman, W. W., Hutchinson, A., Smallwood, P., Nathans, J., Rogan, P. K., Schneider, T. D., & Dean, M. (1998). Organization of the ABCR gene: analysis of promoter and splice junction sequences. Gene, 215, 111–122.

    Article  PubMed  CAS  Google Scholar 

  3. Doyle, L. A., Yang, W., Abruzzo, L. V., Krogmann, T., Gao, Y., Rishi, A. K., & Ross, D. D. (1998). A multidrug resistance transporter from human MCF-7 breast cancer cells. Proceedings of the National Academy of Sciences of United States of America, 95, 15665–15670.

    Article  CAS  Google Scholar 

  4. Miyake, K., Mickley, L., Litman, T., Zhan, Z., Robey, R., Cristensen, B., Brangi, M., Greenberger, L., Dean, M., Fojo, T., & Bates, S. E. (1999). Molecular cloning of cDNAs which are highly over expressed in mitoxantrone-resistant cells: Demonstration of homology to ABC transport genes. Cancer Research, 59, 8–13.

    PubMed  CAS  Google Scholar 

  5. van Herwaarden, A. E., Wagenaar, E., Karnekamp, B., Merino, G., Jonker, J. W., & Schinkel, A. H. (2006). Breast cancer resistance protein (Bcrp1/Abcg2) reduces systemic exposure of the dietary carcinogens aflatoxin B1, IQ and Trp-P-1 but also mediates their secretion into breast milk. Carcinogenesis, 27(1), 123–130.

    Article  PubMed  Google Scholar 

  6. Tantia, M. S., Vijh, R. K., Mishra, B. P., Mishra, B., Kumar, S. B., & Sodhi, M. (2006). DGATI and ABCG2 polymorphism in India cattle (Bos indicus) and buffalo (Bubalus bubalis) breeds. BMC Veterinary Research, 2, 32.

    Article  PubMed  Google Scholar 

  7. van Herwaarden, A. E., Wagenaar, E., Merino, G., Jonker, J. W., Rosing, H., Beijnen, J. H., & Schinkel, A. H. (2007). Multidrug transporter ABCG2/Breast cancer resistance protein secretes riboflavin (vitamin B2) into milk. Molecular and Cellular Biology, 27(4), 1247–1253.

    Article  PubMed  Google Scholar 

  8. Cohen-Zinger, M., Seroussi, E., Larkin, D. M., Loor, J. J., der Wind, A. E., Lee, J. -H., Drackley, J. K., Band, M. R., Hernandez, A. G., Shani, M., Lewin, H. A., Weller, J. I., & 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 Hostein cattle. Genome Research, 15 , 936–944.

    Article  Google Scholar 

  9. Jonker, J. W., Merino, G., Musters, S., van Herwaarden, A. E., Bolscher, E., Wagenaar, E., Mesman, E., Dale, T. C., & Schinkel, A. H. (2005). The breast cancer resistance protein BCRP (ABCG2) concentrates drugs and carcinogenic xenotoxins into milk. Nature Medicine, 11 , 127–129.

    Article  PubMed  CAS  Google Scholar 

  10. Szatmari, I., Vámosi G, Brazda, P., Balint, B. L., Benko, S., Széles L, Jeney, V., Özvegy-Laczka, C., Szanto, A., Barta, E., Balla, J., Sarkadi, B., & Nagy, L. (2006). PPAR regulated ABCG2 expression confers cytoprotection to human dendritic cells. Journal of Biological Chemistry, 281, 23812–23823.

    Article  PubMed  CAS  Google Scholar 

  11. Takenaka, K., Morgan, J. A., Scheffer, G. L., Adachi, M., Stewart, C. F., Sun, D., Leggas, M., Ejendal, K. F., Hrycyna, C. A., & Schuetz, J. D. (2007). Substrate overlap between Mrp4 and Abcg2/Bcrp affects purine analogue drug cytotoxicity and tissue distribution. Cancer Research, 67(14), 6965–6972.

    Article  PubMed  CAS  Google Scholar 

  12. Hori, S., Ohtsuki, S., Tachikawa, M., Kimura, N., Kondo, T., Watanabe, M., Nakashima, E., Terasaki, T. (2004). Functional expression of rat ABCG2 on the luminal side of brain capillaries and its enhancement by astrocyte-derived soluble factor(s). Journal of Neurochemistry, 90(3), 526–536.

    Article  PubMed  CAS  Google Scholar 

  13. Pulido, M. M., Molina, A. J., Merino, G., Mendoza, G., Prieto, J. G., & Alvarez, A. I. (2006). Interaction of enrofloxacin with breast cancer resistance protein (BCRP/ABCG2): Influence of flavonoids and role in milk secretion in sheep. Journal of Veterinary Pharmacology and Therapeutics, 29(4), 279–287.

    Article  PubMed  CAS  Google Scholar 

  14. Ueda, T., Brenner, S., Malech, H. L., Langemeijer, S. M., Perl, S., Kirby, M., Phang, O. A., Krouse, A. E., Donahue, R. E., Kang, E. M., & Tisdale, J. F. (2005). Cloning and functional analysis of the rhesus macaque ABCG2 gene: Forced expression confers an SP phenotype among hematopoietic stem cell progeny in vivo. Journal of Biological Chemistry, 280(2), 991–998.

    Article  PubMed  CAS  Google Scholar 

  15. Eisenblatter, T., & Galla, H. J. (2002). A new multidrug resistance protein at the blood-brain barrier. Biochemical and Biophysical Research Communications, 293(4), 1273–1278.

    Article  PubMed  Google Scholar 

  16. Sanchez-Fernandez, R., Davies, T. G. E., Coleman, J. O. D., & Rea, P. A. (2001). The Arabidopsis thaliana ABC protein superfamily, a complete inventory. The Journal of Biological Chemistry, 276, 30231–30244.

    Article  PubMed  CAS  Google Scholar 

  17. Otsu, C. T., da Silva, I., de Molfetta, J. B., da Silva, L. R., de Almeida-Engler, J., Engler, G., Torraca, P. C., Goldman, G. H., & Goldman, M. H. (2004). NtWBC1, an ABC transporter gene specifically expressed in tobacco reproductive organs. Journal of Experimental Botany, 55(403), 1643–1654.

    Article  PubMed  CAS  Google Scholar 

  18. Hofmann, K., & Stoffel, W. (1993). TMbase-a database of membrane spanning proteins segments. Biological Chemistry Hoppe-Seyler, 374, 166.

    Google Scholar 

  19. Henriksen, U., Fog, J. U., Litman, T., & Gether, U. (2005). Identification of intra-and intermolecular disulfide bridges in the multidrug resistance transporter ABCG2. Journal of Biological Chemistry, 280(44), 36926–36934.

    Article  PubMed  CAS  Google Scholar 

  20. Bhatia, A., Schafer, H. -J., & Hrycyna, C. A. (2005). Oligomerization of the human ABC transporter ABCG2: Evaluation of the naïve protein and chimeric dimers. Biochemistry, 44(32), 10893–10904.

    Article  PubMed  CAS  Google Scholar 

  21. Polgar, O., Robey, R. W., Morisaki, K., Dean, M., Michejda, C., Sauna, Z. E., Ambudkar, S. V., Tarasova, N., & Bates, S. E. (2004). Mutational analysis of ABCG2: Role of the GXXXG motif. Biochemistry, 43(29), 9448–9456.

    Article  PubMed  CAS  Google Scholar 

  22. Zhou, N. E., Kay, C. M., & Hodges, R. S. (1992). Synthetic model proteins. Positional effects of interchain hydrophobic interactions on stability of two-stranded alpha-helical coiled-coils. Journal of Biological Chemistry, 267(4), 2664–2670.

    PubMed  CAS  Google Scholar 

  23. Konig, P., & Richmond, T. J. (1993). The X-ray structure of the GCN4-bZIP bound to ATF/CREB site DNA shows the complex depends on DNA flexibility. Journal of Molecular Biology, 233(1), 139–154.

    Article  PubMed  CAS  Google Scholar 

  24. Epstein, H., & Fischman, D. (1991). Molecular analysis of protein assembly in muscle development. Science, 251, 1039–1044.

    Article  PubMed  CAS  Google Scholar 

  25. Gentz, R., Rauscher F. J. III, Abate, C., & Curran, T. (1989). Parallel association of Fos and Jun leucine zippers juxtaposes DNA binding domains. Science, 243, 1695–1699.

    Article  PubMed  CAS  Google Scholar 

  26. Flemington, E., Speck, S. H. (1990). Evidence for coiled-coil dimer formation by an Epstein-Barr virus transactivator that lacks a heptad repeat of leucine residues. Proceedings of the National Academy of Science of the United States of America, 87, 9459–9463.

    Article  CAS  Google Scholar 

  27. Schneider, E., & Hunke, S. (1998). ATP-binding-cassette (ABC) transport systems: Functional and structural aspects of the ATP-hydrolyzing subunits/domains. FEMS Microbiology Reviews, 22, 1–20.

    Article  PubMed  CAS  Google Scholar 

  28. Higgins, C. F. (2001). ABC transporters: Physiology, structure and mechanism—an overview. Research in Microbiology, 152(3–4), 205–210.

    Article  PubMed  CAS  Google Scholar 

  29. Ambudkar, S. V., Dey, S., Hrycyna, C. A., Ramachandra, M., Pastan, I., & Gottesman, M. M. (1999). Biochemical, cellular, and pharmacological aspects of the multidrug transporter. Annual Review of Pharmacology and Toxicology, 39, 361–398.

    Article  PubMed  CAS  Google Scholar 

  30. Merino, G., Jonker, J. W., Wagenaar, E., van Herwaarden, A. E., & Schinkel, A. H. (2005). The breast cancer resistance protein (BCRP/ABCG2) affects pharmacokinetics, hepatobiliary excretion, and milk secretion of the antibiotic nitrofurantoin. Molecular Pharmacology, 67, 1758–1764.

    Article  PubMed  CAS  Google Scholar 

  31. Merino, G., Álvarez AI, Pulido, M. M., Molina, A. J., Schinkel, A. H., & Prieto, J. G. (2006). Breast cancer resistance protein (BCRP/ABCG2) transports fluoroquinolone antibiotics and affects their oral availability, pharmacokinetics, and milk secretion. Drug Metabolism and Disposition Fast Forward, 34, 690–695.

    Article  CAS  Google Scholar 

  32. Miri, C. Z., Seroussi, E., Larkin, D. M., Loor, J., Wind, A. E., Lee, J. H., Drackley, J. K., Band, M., & Hernandez, A. G. (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 Research, 15, 936–944.

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by the Program for New Century Excellent Talents in Chinese Universities (NO. NCET-05-0857) and Post-Graduate Innovation Program of Northwest A&F University (NO. 05YCH021), in China. Sincere gratitude is also expressed to Dr. George Acquaah, Chairman of the Department of Agriculture and Natural Resources at Langston University, for editorial assistance.

Author information

Authors and Affiliations

  1. College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, 712100, P.R. China

    Hui Juan Wu, Jun Luo, Li Juan Zhang, Xue Feng Han & Bao Jin Yang

  2. Center for Biotechnology Research and Education, Langston University, Langston, OK, 73050, USA

    Ning Wu & Kanyand Matand

Authors
  1. Hui Juan Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jun Luo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ning Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kanyand Matand
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Li Juan Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xue Feng Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Bao Jin Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jun Luo.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wu, H.J., Luo, J., Wu, N. et al. Cloning, Sequence and Functional Analysis of Goat ATP-binding Cassette Transporter G2 (ABCG2). Mol Biotechnol 39, 21–27 (2008). https://doi.org/10.1007/s12033-007-9024-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12033-007-9024-5

Keywords

Search

Navigation