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Cloning and Expression of Bovine Glucose Transporter GLUT12

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Abstract

GLUT12 is a new member of facilitative glucose transporters. It was originally cloned from a human breast cancer cell line and its expression has been detected in rat mammary gland. Glucose transport across the plasma membrane of mammary epithelial cells is a rate-limiting factor in milk production. To examine GLUT12’s expression and facilitate the study of GLUT12’s potential role in supporting milk synthesis in lactating bovine mammary gland, we cloned bovine GLUT12 and examined its distribution of mRNA expression in bovine tissues. The full-length mRNA of bGLUT12 is 2423 base pairs long and is predicted to encode a protein of 621 amino acids with a molecular weight of approximately 67 kDa. The deduced amino acid sequence of bovine GLUT12 is 87% and 82% identical to the sequences of human and mouse GLUT12. The sequence of bGLUT12 contains several characteristically conserved sugar transporter family signatures. Analysis of current bovine genomic data indicates that bovine GLUT12 gene consists of five exons. The major in vitro transcription and translation product of bovine GLUT12 cDNA migrated at an apparent molecular weight of 41 kDa. In the presence of canine microsomal membranes, the translation product increased to 43 kDa, suggesting glycosylation. GLUT12 mRNA was found in all bovine tissues examined, but most abundant in bovine spleen and skeletal muscle, at intermediate levels in bovine kidney, testes, and mammary gland, and at lower levels in bovine liver, lung and intestine. Immunofluorescence staining showed that, in the presence of insulin, bGLUT12 is mainly distributed in the cytoplasm of the transiently transfected MAC-T bovine mammary epithelial cells.

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Abbreviations

bGLUT12 = :

bovine solute carrier family 2 member 12 (gene symbol SLC2A12),

EST = :

expressed sequence tag,

kb = :

kilobases,

poly(A)+ = :

polyadenylated RNA,

RACE = :

rapid amplification of cDNA ends,

RT-PCR = :

reverse transcription-polymerase chain reaction,

TM = :

transmembrane domain,

UTR = :

untranslated region.

References

  • Ahmed N, Berridge MV (1999) N-glycosylation of glucose transporter-1 (Glut-1) is associated with increased transporter affinity for glucose in human leukemic cells. Leuk Res 23: 395–401

    PubMed  CAS  Google Scholar 

  • Bell AW, Bauman DE (1997) Adaptations of glucose metabolism during pregnancy and lactation. J Mammary Gland Biol Neoplasia 2: 265–278

    Article  PubMed  CAS  Google Scholar 

  • Dingwall C, Laskey RA (1991) Nuclear targeting sequences—a consensus. Trends Biochem Sci 16: 478-481

    PubMed  CAS  Google Scholar 

  • Huynh HT, Robitaille G, Turner JD (1991) Establishment of bovine mammary epithelial cells (MAC-T): an in vitro model for bovine lactation. Exp Cell Res 197: 191-199

    Article  PubMed  CAS  Google Scholar 

  • Ibberson M, Uldry M, Thorens B (2000) GLUTX1, a novel mammalian glucose transporter expressed in the central nervous system and insulin-sensitive tissues. J Biol Chem 275: 4607-4612

    Article  PubMed  CAS  Google Scholar 

  • Joost JG, Thorens B (2001) The extended GLUT-family of sugar/polyol transport facilitators: nomenclature, sequence characteristics, and potential function of its novel members (review). Mol Membr Biol 18: 247-256

    PubMed  CAS  Google Scholar 

  • Khan AH, Capilla E, Hou JC, Watson RT, Smith JR, et al. (2004) Entry of newly synthesized GLUT4 into the insulin-responsive storage compartment is dependent upon both the amino terminus and the large cytoplasmic loop. J Biol Chem 279: 37505-37511

    PubMed  CAS  Google Scholar 

  • Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157: 105–132

    Article  PubMed  CAS  Google Scholar 

  • Lisinski I, Schurmann A, Joost HG, Cushman SW, Al-Hasani H (2001) Targeting of GLUT6 (formerly GLUT9) and GLUT8 in rat adipose cells. Biochem J 358: 517–522

    Article  PubMed  CAS  Google Scholar 

  • Macheda ML, Williams ED, Best JD, Wlodek ME, Rogers S (2003) Expression and localisation of GLUT1 and GLUT12 glucose transporters in the pregnant and lactating rat mammary gland. Cell Tissue Res 311: 91–97

    Article  PubMed  CAS  Google Scholar 

  • Mepham TB (1993) The development of ideas on the role of glucose in regulating milk secretion. Aust J Agric Res 44: 509–522

    Article  CAS  Google Scholar 

  • Rogers S, Macheda ML, Docherty SE, Carty MD, Henderson MA, et al. (2002) Identification of a novel glucose transporter-like protein—GLUT-12. Am J Physiol Endocrinol Metab 282: E733–E738

    PubMed  CAS  Google Scholar 

  • Rogers S, Chandler JD, Clarke AL, Petrou S, Best JD (2003) Glucose transporter GLUT12-functional characterization in Xenopus laevis oocytes. Biochem Biophys Res Commun 308: 422–426

    Article  PubMed  CAS  Google Scholar 

  • Samih N, Hovsepian S, Aouani A, Lombardo D, Fayet G (2000) Glut-1 translocation in FRTL-5 thyroid cells: role of phosphatidylinositol 3-kinase and N-glycosylation. Endocrinology 41: 4146–4155

    Google Scholar 

  • Samih N, Hovsepian S, Notel F, Prorok M, Zattara-Cannoni H, et al. (2003) The impact of N- and O-glycosylation on the functions of Glut-1 transporter in human thyroid anaplastic cells. Biochim Biophys Acta 1621: 92–101

    PubMed  CAS  Google Scholar 

  • Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucleic Acids Res 22: 4673–4680

    PubMed  CAS  Google Scholar 

  • Uldry M, Ibberson M, Horisberger JD, Chatton JY, Riederer BM, et al. (2001) Identification of a mammalian H(+)-myo-inositol symporter expressed predominantly in the brain. EMBO J 20: 4467–4477

    Article  PubMed  CAS  Google Scholar 

  • Watson RT, Kanzaki M, Pessin JE (2004) Regulated membrane trafficking of the insulin-responsive glucose transporter 4 in adipocytes. Endocr Rev 25: 177-204

    Article  PubMed  CAS  Google Scholar 

  • Zhao F-Q, Miller PJ, Wall EH, Zheng Y-C, Dong B, et al. (2004) Bovine glucose transporter GLUT8: cloning, expression, and developmental regulation in mammary gland. Biochim Biophys Acta 1680: 103–113

    PubMed  CAS  Google Scholar 

  • Zhao F-Q, Zheng Y-C, Wall EH, McFadden TB (2005) Cloning and Expression of Bovine sodium/Glucose Cotransporters. J Dairy Sci 88: 182–194

    PubMed  CAS  Google Scholar 

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Acknowledgments

This project was supported by National Research Initiative Competitive Grant No. 2005-35206-15267 from the USDA Cooperative State Research, Education, and Extension Service (to FQZ).

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Authors and Affiliations

  1. Lactation and Mammary Gland Biology Group, Department of Animal Science, University of Vermont, RM 219 Terrill Building,570 Main Street, Burlington, Vermont, 05405, USA

    Peter J. Miller, Kiera A. Finucane, Megan Hughes & Feng-Qi Zhao

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  1. Peter J. Miller
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  2. Kiera A. Finucane
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  3. Megan Hughes
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  4. Feng-Qi Zhao
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Corresponding author

Correspondence to Feng-Qi Zhao.

Additional information

The nucleotide sequence data reported in this paper have been submitted to GenBank with assigned accession number AY514443.

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Miller, P.J., Finucane, K.A., Hughes, M. et al. Cloning and Expression of Bovine Glucose Transporter GLUT12. Mamm Genome 16, 873–883 (2005). https://doi.org/10.1007/s00335-005-0080-5

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  • DOI: https://doi.org/10.1007/s00335-005-0080-5

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