The RFA regulatory sequence-binding protein in the promoter of prostate-specific antigen gene

  • Weiwen Chen
  • Jianye Zhang
  • Charles Y. F. Young
  • Lianying Zhang
  • Liucun Chen
  • Jian Zhao
Article
  • 14 Downloads

Abstract

To assure what sequence associated with the androgen regulation, a 15 bp region at the upstream of the ARE of prostate-specific antigen (PSA) promoter, termed RFA, was found indispensable for androgen receptor (AR)-mediated transactivation of PSA promoter. In transfection and CAT assays, some nucleotides substitution in RFA could significantly decrease the androgen inducibility for PSA promoter. The in vitro DNA binding assay demonstrated that RFA bound specifically with some non-receptor protein factors in prostate cell nucleus, but the mutant type of RFA lost this ability, so RFA might be a novel accessory cis-element. The RFA-binding proteins were isolated and purified by affinity chromatography using RFA probes. SDS-PAGE and preliminary protein identification showed these proteins possessed sequence high homology with multifunctional protein heterogeneous nuclear ribonucleoprotein A1, A2 (hnRNP A1, A2). RFA-binding proteins possibly cooperate with AR-mediated transactivation for PSA promoter as coactivator. The study results will facilitate further understanding the mechanism and tissue specificity of PSA promoter.

Keywords

prostate-specific antigen promoter heterogeneous nuclear ribonucleoprotein A1 A2 

References

  1. 1.
    Woodward, R. N., Li, M., Holland, L. J., Novel accessory factor-binding site required for Glucocorticoid regulation of the gamma-fibrinogen subunit gene fromxerropus Laevis, Mol. Endocrinol., 1997, 11(5): 563–576.PubMedCrossRefGoogle Scholar
  2. 2.
    Ho, K. C., Marche, K. B., Tan, J. A. et al., A complex response element in intron 1 of the androgen-regulated 20-kDa protein gene displays cell type-dependent androgen receptor specificity, J. Biol. Chem., 1993, 268: 27226–27235.PubMedGoogle Scholar
  3. 3.
    Riegman, P. H., Vlietstra, R. J., van der Korput, J. A. et al., The promoter of the prostate-specific antigen gene contains a functional androgen responsive element, Mol. Endocrinol., 1991, 5: 1921–1930.PubMedCrossRefGoogle Scholar
  4. 4.
    Zhang, J. Y., Pang, W. Q., Zhang, L. Y. et al., Primary analysis of a regulation sequence in the promoter region of human prostate specific antigen gene, Chin. J. Med. Genet. (in Chinese), 1997, 14(1): 24–27.Google Scholar
  5. 5.
    Zhang, J. Y., Pang, W. Q., Wang, X. et al., A sequence associated with the regulation of androgen in PSA gene promoter, Prog. Biochem. Biophys. (in Chinese), 1997, 24: 228–232.Google Scholar
  6. 6.
    Bagchi, M. K., Tsai, S. Y., Tsai, M. J. et al., Identification of a functional intermediate in receptor activation in progesterone-dependent cell-free transcription, Nature, 1990, 345: 547–550.PubMedCrossRefGoogle Scholar
  7. 7.
    Murtha, P., Tindall, D. J., Young, C. Y. F., Androgen induction of a human prostate-specific kallikrein, hKLK2: characterization of an androgen response element in the 5′ promoter region of the gene, Biochemistry, 1993, 32(25): 6459–6464.PubMedCrossRefGoogle Scholar
  8. 8.
    Renie, P. S., Bruchovsky, N., Leco, K. J., et al., Characterization of two cis-acting DNA elements involved in the androgen regulation of the probasin gene, Mol. Endocrinol., 1993, 7: 3–26.CrossRefGoogle Scholar
  9. 9.
    Marshak, D. R., Kadonaga, T. K., Burgess, R. R. et al., Strategies for Protein Purification and Characterization (in Chinese, translated by Zhu Houchu), Beijing: Science Press, 1997, 101–117.Google Scholar
  10. 10.
    Tomonaga, T., Levens, D., Heterogeneous nuclear ribonucleoprotein K is a DNA-binding transactivator, J. Biol. Chem., 1995, 270:4875–4881.PubMedCrossRefGoogle Scholar
  11. 11.
    Miau, L. H., Chang, C. J., Shen, B. J. et al., Identification of heterogeneous nuclear ribonucleoprotein K (hnRNP K) as a repressor of C/EBP beta-mediated gene activation, J. Biol. Chem., 1998, 273: 10784–10791.PubMedCrossRefGoogle Scholar
  12. 12.
    Chen, H., Hu, B., Allegretto, E. A. et al., The vitamin D response element-binding protein—A novel dominant-negative regulator of vitamind-directed transactivation, J. Biol. Chem., 2000, 275(45): 35557–35564.PubMedCrossRefGoogle Scholar
  13. 13.
    Leverrier, S., Cinato, E., Paul, C. et al., Purification and cloning of type A/B hnRNP proteins involved in transcriptional activation from the Rat spi 2 gene GAGA box, Biol. Chem., 2000, 381(11): 1031–1040.PubMedCrossRefGoogle Scholar
  14. 14.
    Kamma, H., Fujimoto, M., Fujiwara, M. et al., Interaction of hnRNP A2/B1 isoforms with telomeric ssDNA and the in vitro function, Biochem. Biophys. Res. Commun., 2001, 280(3): 625–630.PubMedCrossRefGoogle Scholar
  15. 15.
    Man, Y. G., Martinez, A., Avis, I. M. et al., Phenotypically different cells with heterogeneous nuclear ribonucloprotein A2/B1 overexpression show similar genetic alterations, Am. J. Respir. Cell Mol. Biol., 2000, 23(5): 636–645.PubMedGoogle Scholar

Copyright information

© Science in China Press 2003

Authors and Affiliations

  • Weiwen Chen
    • 1
  • Jianye Zhang
    • 1
  • Charles Y. F. Young
    • 2
  • Lianying Zhang
    • 1
  • Liucun Chen
    • 1
  • Jian Zhao
    • 3
  1. 1.Institute of Biochemistry and Molecular Biology, School of MedicineShandong UniversityJinanChina
  2. 2.Department of Urology Research of Mayo Clinic/FoundationUSA
  3. 3.Department of Thoracic SurgeryQilu Hospital of Shandong UniversityJinanChina

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