Identification of a short PIASx gene promoter that directs male germ cell-specific transcription in vivo

• Published in: Biochemical and biophysical research communications 2003-08, Vol.308 (1), p.139-147
• Main Authors: Santti, Henrikki, Mikkonen, Laura, Hirvonen-Santti, Sirpa, Toppari, Jorma, Jänne, Olli A, Palvimo, Jorma J
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Carrier Proteins - chemistry; Carrier Proteins - genetics; DNA; Electrophoretic Mobility Shift Assay; GC box; Genes, Reporter; Germ Cells - metabolism; Ligases - chemistry; Ligases - genetics; Male; Male germ cell; Mice; Mice, Transgenic; Molecular Sequence Data; PIASx; Promoter Regions, Genetic; Protein Binding; Protein Inhibitors of Activated STAT; Sequence Analysis, DNA; Small Ubiquitin-Related Modifier Proteins; Spermatogenesis; Spermatozoa - metabolism; Sumoylation; Testis; Transcription; Transcription Factors - metabolism; Transcription, Genetic - genetics; Ubiquitin-Protein Ligases
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/S0006-291X(03)01339-1

PIASx gene encodes two SUMO E3 ligases that are highly expressed in the testis. We have isolated and analyzed the promoter of the murine PIASx gene. Electrophoretic mobility shift assays with testicular nuclear extracts showed that the proximal promoter forms a major DNA–protein complex containing Sp1, Sp2, and Sp3 transcription factors. Reporter gene assays in cultured cells indicated that a fragment comprising nucleotides from −168 to +76 relative to transcription start site is sufficient for basal promoter activity in cultured cells, but these in vitro assays failed to reveal clear differences in promoter activity between testis- and non-testis-derived cell lines. Interestingly, however, the proximal promoter encompasses the elements necessary for a testis-specific transcription in vivo, as it directed β-galactosidase expression exclusively to male germ cells in transgenic mice. In conclusion, we have characterized the minimal PIASx promoter that can be used for highly specific targeting of transgene expression to male germ cells.