Spermatogenesis-related ring finger gene ZNF230 promoter: identification and functional analysis

Mol Biol Rep. 2009 May;36(5):1187-93. doi: 10.1007/s11033-008-9296-2. Epub 2008 Jun 27.

Abstract

The ZNF230 gene is a recently cloned gene which is transcribed only in fertile male testes and may be related to human spermatogenesis. To characterize the multiple stage-specific transcription elements necessary for ZNF230 expression, we cloned ZNF230 promoter and constructed chimeric luciferase reporter Plasmids. Overexpression and site-directed mutation test were used to characterize the cis-element. The results showed ZNF230 gene promoter to be GC rich and not contain a TATA box. Deletion analysis of the 5'-flanking region of ZNF230 in HEK293 cells indicated that the sequence encompassing from nt -131 to +152 has a basal transcriptional activity. Site-directed mutation test and mithramycin A treatment demonstrated that the ZNF230 promoter contained a functional Sp1 site. Overexpression of the Sox5 protein activated the promoter activity. A 312-bp fragment surrounding the transcription start site exhibits a characteristic CpG island which overlaps with the promoter region. We also provided evidence that both the human and mouse znf230 promoter consist of Sp1 binding site and GC-rich sequences, suggesting Sp1 is required for the transcription of human and mouse ZNF230 genes. In conclusion, these findings suggest that ZNF230 is tightly controlled at transcriptional level and a common mechanism controls the basal transcription of ZNF230 gene.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites
  • CpG Islands / genetics
  • DNA-Binding Proteins / genetics*
  • Gene Expression Regulation
  • Humans
  • Male
  • Mice
  • Molecular Sequence Data
  • Organ Specificity
  • Promoter Regions, Genetic*
  • SOXD Transcription Factors / metabolism
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Sequence Deletion
  • Sequence Homology, Nucleic Acid
  • Sp1 Transcription Factor / metabolism
  • Spermatogenesis / genetics*
  • Spermatozoa / metabolism
  • Transcription Factors / genetics*
  • Transcription, Genetic

Substances

  • DNA-Binding Proteins
  • SOX5 protein, human
  • SOXD Transcription Factors
  • Sp1 Transcription Factor
  • Transcription Factors
  • ZNF230 protein, human