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Molecular cloning and structural characterization of the functional human thymosin β4 gene

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Abstract

Thymosin β4 (Tβ4), the actin-sequestering protein that regulates the polymerization and depolymerization of actins, is widely distributed in animal tissues. Expression of this gene in human cells is developmentally regulated and appears to play critical roles in tumorigenesis. As a first step toward analyzing the transcriptional regulation of human Tβ4 (hTβ4), we isolated the gene encoding this peptide and characterized its structure features. Human Tβ4 gene comprises three exons and two introns distributed over 2 kb with its transcription start site at 72 bp upstream of the initiation codon. Expression of the chloramphenicol acetyltransferase (CAT) reporter constructs directed by various parts of the 5′-flanking region of this gene was evaluated by transient transfection assays using human colorectal carcinoma SW480 cells as hosts. Significant promoter activity was found in the −437 to +29 region of hTβ4 gene even though it lacks both TATA and CCAAT boxes. However, cis-element(s) responsible for PMA-induced expression of Tβ4 gene was not identified within its 1.5-kb 5′-flanking region. Taken together, our data provide crucial information for further dissection of the molecular mechanism(s) underlying aberrant expression of the Tβ4 gene during malignant progression of human cancers.

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References

  1. Goldstein AL, Slater FD, White A: Preparation, assay and partial purification of a thymic lymphocytopoietic factor (thymosin). Proc Natl Acad Sci USA 56: 1010–1017, 1966

    CAS  PubMed  Google Scholar 

  2. Huff T, Muller CSG, Otto AM, Netzker R, Hannappel E: β-thymosin, small acidic peptides with multiple functions. Int J Biochem Cell Biol 33: 205–220, 2001

    Article  CAS  PubMed  Google Scholar 

  3. Goldstein AL, Guha A, Zatz MM, Hardy MA, White A: Purification and biological activity of thymosin, a hormone of the thymus gland. Proc Natl Acad Sci USA 69: 1800–1803, 1972

    CAS  PubMed  Google Scholar 

  4. Safer D, Elzinga M, Nachmias VT: Thymosin beta 4 and Fx, an actin-sequestering peptide, are indistinguishable. J Biol Chem ([0-9]+): 4029–4032, 1991

    CAS  PubMed  Google Scholar 

  5. Leonard DGB, Ziff EB, Greene LA: Identification and characterization of mRNAs regulated by nerve growth factor in PC12 cells. Mol Cell Biol 7: 3156–3167, 1987

    CAS  PubMed  Google Scholar 

  6. Shimamura R, Kudo J, Kondo H, Dohmen K, Gondo H, Okamura S, Ishibashi H, Niho Y: Expression of the thymosin beta 4 gene during differentiation of hematopoietic cells. Blood 76: 977–984, 1990

    CAS  PubMed  Google Scholar 

  7. Grant DS, Kinsella JL, Kibbey MC, LaFlamme S, Burbelo PD, Goldstein AL, Kleinman HK: Matrigel induces thymosin beta 4 gene in differentiating endothelial cells. J Cell Sci 108: 3685–3694, 1995

    CAS  PubMed  Google Scholar 

  8. Oh IS, So SS, Jahng KY, Kim HG: Hepatocyte growth factor upregulates thymosin beta 4 in human umbilical vein endothelial cells. Biochem Biophys Res Commun 296: 401–405, 2002

    CAS  PubMed  Google Scholar 

  9. Li QY, Jones PL, Lafferty RP, Safer D, Levy RJ: Thymosin beta 4 regulation, expression and function in aortic valve interstitial cells. J Heart Valve Dis 11: 726–735, 2002

    PubMed  Google Scholar 

  10. Blain EJ, Mason DJ, Duance VC: The effect of cyclical compressive loading on gene expression in articular cartilage. Biorheology 40: 111–117, 2003

    CAS  PubMed  Google Scholar 

  11. Golla R, Philp N, Safer D, Chintapalli J, Hoffman R, Collins L, Nachmias VT:. Coordinate regulation of the cytoskeleton in 3T3 cells overexpressing thymosin-beta4. Cell Motil Cytoskeleton 38: 187–200, 1997

    CAS  PubMed  Google Scholar 

  12. Conlon JM, Grimelius L, Wallin G., Thim L: Isolation and structural characterization of thymosin beta 4 from human medullary thyroid carcinoma. J Endocrinol 118: 155–159, 1988

    CAS  PubMed  Google Scholar 

  13. Hall AK: Differential expression of thymosin genes in human tumors and in the developing human kidney. Int J Cancer 48: 672–677, 1991

    CAS  PubMed  Google Scholar 

  14. Santelli G, Califano D, Chiappetta G, Vento MT, Bartoli PC, Zullo F, Trapasso F, Viglietto G, Fusco A: Thymosin beta-10 gene overexpression is a general event in human carcinogenesis. Am J Pathol 155: 799–804, 1999

    CAS  PubMed  Google Scholar 

  15. Xie D, Jauch A, Miller CW, Bartram CR, Koeffler HP: 2002. Discovery of over-expressed genes and genetic alterations in breast cancer cells using a combination of suppression subtractive hybridization, multiplex FISH and comparative genomic hybridization. Int J Oncol 21: 499–507, 2002

    CAS  PubMed  Google Scholar 

  16. Clark EA, Golub TR, Lander ES, Hynes RO: Genomic anaylasis of metastasis reveals and essential role for RhoC. Nature 406: 532–535, 2000

    CAS  PubMed  Google Scholar 

  17. Kobayashi T, Okada F, Fujii N, Tomita N, Ito S, Tazawa H, Aoyama T, Choi SK, Shibata T, Fujita H, Hosokawa M: Thymosin-beta4 regulates motility and metastasis of malignant mouse fibrosarcoma cells. Am J Pathol 160: 869–882, 2002

    CAS  PubMed  Google Scholar 

  18. Wang WS, Chen PM, Hsiao HL, Ju SY, Su Y. Overexpression of the thymosin β-4 gene is associated with malignant progression of SW480 colon cancer cells. Oncogene 22: 3297–3306, 2003

    CAS  PubMed  Google Scholar 

  19. Wang WS, Chen PM, Hsiao HL, Wang HS, Liang WH, Su Y. Overexpression of the thymosin β-4 gene is associated with increased invasion of SW480 colon carcinoma cells and distant metastasis of human colorectal carcinoma. Oncogene 23: 6666–6671, 2004

    CAS  PubMed  Google Scholar 

  20. Varghese S, Kronenberg HM: Rat thymosin beta 4 gene. Intron-containing gene and multiple retroposons. J Biol Chem 266: 14256–14261, 1991

    CAS  PubMed  Google Scholar 

  21. Li X, Zimmerman A, Copeland NG, Gilbert DJ, Jenkins NA, Yin HL: The mouse thymosin beta 4 gene: structure, promoter identification, and chromosome localization. Genomics 32: 388–394, 1996

    CAS  PubMed  Google Scholar 

  22. Su Y, Chang SL, Hsiao HL: Characterization of a 5′-flanking region supporting the transcription of mouse thymosin β-4 in mouse NIH3T3 cells. Mol Cell Biochem 203: 163–167, 2000

    CAS  PubMed  Google Scholar 

  23. Clauss LM, Wathelet MG, Szpirer J, Islam MQ, Levan G, Szpirer C, Huez, GA: Human thymosin-β4/6–26 gene is part of a multigene family composed of seven members located on seven different chromosomes. Genomics 9: 174–180, 1991

    CAS  PubMed  Google Scholar 

  24. Lahn BT, Page DT: Functional coherence of the human Y chromosome. Science 278: 675–680, 1997

    CAS  PubMed  Google Scholar 

  25. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. 1989

    Google Scholar 

  26. Kanamoto R, Su Y, Pitot HC: Hormonal regulation of serine dehydratase gene expression in liver and kidney of the adrenalectomized rat. Mol Endocrinol 5: 1661–1668, 1991

    CAS  PubMed  Google Scholar 

  27. Tang RY, SuY: Construction of a cell-based high-flux assay for the rev protein of HIV-1. J Virol Methods 65: 153–158, 1997

    CAS  PubMed  Google Scholar 

  28. Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Bio 162: 156–159, 1987

    CAS  Google Scholar 

  29. Breathnach R, Chambon P: Organization and expression of eukaryotic split genes coding for proteins. Annu Rev Biochem 50: 349–383, 1981

    CAS  PubMed  Google Scholar 

  30. Pugh BF, Tjian R: Transcription from a TATA-less promoter requires a multisubunit TFIID complex. Genes Dev 5: 1935–1945, 1991

    CAS  PubMed  Google Scholar 

  31. Lu WC, Liu YN, Kang BB, Chen JH: Trans-activation of heparanase promoter by ETS transcription factors. Oncogene 22: 919–923, 2003

    CAS  PubMed  Google Scholar 

  32. Zhu GH, Lenzi M, Schwartz EL: The Sp1 transcription factor contributes to the tumor necrosis factor-induced expression of the angiogenic factor thymidine phosphorylase in human colon carcinoma cells. Oncogene 21: 8477–8485, 2002

    CAS  PubMed  Google Scholar 

  33. Zeng Q, McCauley LK, Wang CY: Hepatocyte growth factor inhibits anoikis by induction of activator protein 1-dependent cyclooxygenase-2. J Biol Chem 277: 50137–50142, 2002

    CAS  PubMed  Google Scholar 

  34. Goldstein AL: Thymosin beta 4: A new molecular target for antitumor strategies. J Natl Cancer Inst 95:1646–1647, 2003

    CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department and Institute of Pharmacology, School of Medicine, National Yang-Ming University, Shih-Pai, Taipei, Taiwan, Republic of China

    Shu-Ping Yang & Hui-Ju Lee

  2. Graduate Institute of Biopharmaceutical Science, Faculty of Life Science, National Yang-Ming University, Shih-Pai, Taipei, Taiwan, Republic of China

    Yeu Su

  3. Graduate Institute of Biopharmaceutical Science, National Yang-Ming University, Shih-Pai, Taipei, 11221, Taiwan (ROC)

    Yeu Su

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  1. Shu-Ping Yang
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  2. Hui-Ju Lee
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Correspondence to Yeu Su.

Additional information

Sequence data from this article have been deposited with the EMBL/GenBank Data Libraries under Accession No. AJ295158.

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Yang, SP., Lee, HJ. & Su, Y. Molecular cloning and structural characterization of the functional human thymosin β4 gene. Mol Cell Biochem 272, 97–105 (2005). https://doi.org/10.1007/s11010-005-7642-4

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  • DOI: https://doi.org/10.1007/s11010-005-7642-4

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