Location of promoter elements necessary and sufficient to direct testis-specific expression of the Hst70/Hsp70.2 gene.
The rat Hst70 gene and its mouse counterpart Hsp70.2 are expressed specifically in pachytene primary spermatocytes and spermatids. Here we demonstrate that a 165 bp fragment of the Hst70 gene promoter, containing the T1 transcription start site region, entire exon 1 and 42 bp 5' region of the intron, is sufficient to drive testis-specific expression of the chloramphenicol acetyltransferase reporter gene in transgenic mice with the same developmentally regulated pattern as the endogenous Hsp70.2 gene. We show further that high-level tissue-specific gene expression requires additional sequences localized upstream of the T2 transcription start site. Electrophoretic mobility-shift assay analysis revealed that only testes of juvenile rats, when Hst70 gene expression is repressed, contain proteins that specifically bind to the Oct (octamer) sequence localized directly downstream of the T1 site.
Scieglińska
Dorota
D
Department of Tumour Biology, Centre of Oncology, Maria Skłodowska-Curie Memorial Institute, Wybrzeze Armii Krajowej 15, 44-101 Gliwice, Poland.
Vydra
Natallia
N
Krawczyk
Zdzisław
Z
Widłak
Wiesława
W
D000818:Animals; D001483:Base Sequence; D002457:Cell Extracts; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D005091:Exons / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D007438:Introns / Q000235:genetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D050811:Octamer Transcription Factor-1; D009928:Organ Specificity; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D020218:Response Elements / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016329:Sp1 Transcription Factor / Q000378:metabolism; D013737:Testis / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism; D024363:Transcription Initiation Site; D019076:Transgenes / Q000235:genetics
D002457:Cell Extracts; D004268:DNA-Binding Proteins; D018840:HSP70 Heat-Shock Proteins; C490434:Hspa2 protein, mouse; C490435:Hspa2 protein, rat; D050811:Octamer Transcription Factor-1; C494166:Pou2f1 protein, mouse; C494168:Pou2f1 protein, rat; D012333:RNA, Messenger; D016329:Sp1 Transcription Factor; D014157:Transcription Factors