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Molecular and Cellular Biochemistry

, Volume 276, Issue 1–2, pp 159–167 | Cite as

Characterization of a human TSPY promoter

  • Britta Skawran
  • Stephanie Schubert
  • Frank Dechend
  • Jörg Vervoorts
  • Karim Nayernia
  • Bernhard Lüscher
  • Jörg Schmidtke
Article

Abstract

Human TSPY is a candidate oncogene and is supposed to function as a proliferation factor during spermatogenesis. It is the only mammalian protein-coding gene known to be organized as a tandem repeat gene family. It is expressed at highest level in spermatogonia and to a lower amount in primary spermatocytes. To characterize the human TSPY promoter we used the luciferase reporter system in a mouse spermatogonia derived cell line (GC-1spg) and in a GC-4spc cell line, that harbour prophase spermatocytes of the preleptotene and early pachytene stage. We isolated a 1303 bp fragment of the 5′-flanking region of exon 1 that shows significant promoter activity in GC-1spg and reduced activity in GC-4spc cells. In order to gain further insight into the organization of the TSPY-promoter, stepwise truncations of the putative promoter sequence were performed. The resulting fragments were cloned into the pGL3-vector and analysed for reporter gene activity in the murine germ cell lines GC-1spg and GC-4spc, leading to the characterization of a core promoter (−159 to −1), an enhancing region (−673 to −364) and a silencing region (−1262 to −669). Database research for cis-active elements yielded two putative SOX-like binding sites in the enhancing region and reporter gene activity was drastically reduced when three nucleotides of the AACAAT SOX core sequence were mutated. Our findings strongly suggest that testis-specific expression of human TSPY is mediated by Sox proteins. (Mol Cell Biochem 276: 159–167, 2005)

Keywords

TSPY SOX promoter luciferase regulation testis 

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Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • Britta Skawran
    • 1
    • 6
  • Stephanie Schubert
    • 1
  • Frank Dechend
    • 2
  • Jörg Vervoorts
    • 3
  • Karim Nayernia
    • 4
  • Bernhard Lüscher
    • 5
  • Jörg Schmidtke
    • 1
  1. 1.Institute of Human GeneticsHannover Medical SchoolHannoverGermany
  2. 2.Centre of Reproduction and Human GeneticsLaboratory of Cytogenetic and Molecular GeneticsHildesheimGermany
  3. 3.Institute of Molecular BiologyHannover Medical SchoolHannoverGermany
  4. 4.Institute of Human GeneticsUniversity of GöttingenGöttingenGermany
  5. 5.Department of Biochemistry and Molecular BiologyInstitute of BiochemistryAachenGermany
  6. 6.Institute of Cell and Molecular PathologyHannover Medical SchoolHannoverGermany

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