A gene-specific promoter in transgenic mice directs testis-specific demethylation prior to transcriptional activation In vivo.
Transcription of the autosomal phosphoglycerate kinase gene, Pgk-2, is initiated at the onset of meiosis during spermatogenesis in mammals. However, in the mouse, the 5' portion of the endogenous Pgk-2 coding sequence undergoes a specific demethylation event that precedes transcriptional activation by 10-12 days. Here we show that transgenes consisting of the Pgk-2 core promoter ligated to the CAT reporter gene undergo a similar tissue-, stage-, and cell type-specific demethylation in the 5' portion of the CAT coding sequence, whereas transgenes consisting of the CAT reporter sequence alone, or of the CAT sequence ligated to the CpG island-containing transferrin gene promoter, demonstrate different patterns of demethylation. These results indicate that specific promoter sequences can influence the pattern of tissue-specific demethylation within different genes and that a signal for spermatogenic cell-specific demethylation resides within the core promoter of the mammalian Pgk-2 gene.
Zhang
L P
LP
Department of Genetics, Southwest Foundation for Biomedical Research, San Antonio, Texas 78245, USA.
Stroud
J C
JC
Walter
C A
CA
Adrian
G S
GS
McCarrey
J R
JR
D000818:Animals; D015139:Blotting, Southern; D002415:Cats; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation; D003640:Dealkylation; D005260:Female; D006822:Hybrid Cells; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008822:Mice, Transgenic; D010735:Phosphoglycerate Kinase / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D013091:Spermatogenesis / Q000502:physiology; D013737:Testis / Q000378:metabolism*; D014158:Transcription, Genetic / Q000502:physiology*; D015533:Transcriptional Activation / Q000502:physiology*; D019076:Transgenes / Q000235:genetics
D015500:Chloramphenicol O-Acetyltransferase; D010735:Phosphoglycerate Kinase