Chromosome transfer activates and delineates a locus control region for perforin.
Perforin gene (PRF1) transcription regulates perforin expression in NK cells and CTL. Here we identified the locus-wide ensemble of cis-acting sequences that drives PRF1 transcription physiologically. By using chromosome transfer, we revealed that de novo activation of a silent PRF1 locus was controlled by a 150 kb domain comprised of 16 DNase I hypersensitive sites (DHSs). These cis-acting sequences included a locus control region (LCR) and conferred developmentally appropriate and lineage-specific expression of human perforin from BAC transgenes. The LCR included four distal DHSs that were required for perforin expression from its natural locus, and their engineered deletion from the PRF1 BAC transgene abolished LCR function and led to rapid gene silencing. Thus, LCR function is central for regulating the developmental and activation-specific PRF1 promoter activity characteristic of NK cells and CTL.
Pipkin
Matthew E
ME
Department of Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, FL 33136, USA.
Ljutic
Belma
B
Cruz-Guilloty
Fernando
F
Nouzova
Marcela
M
Rao
Anjana
A
Zúñiga-Pflücker
Juan Carlos
JC
Lichtenheld
Mathias G
MG
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000235:genetics*; D006801:Humans; D017403:In Situ Hybridization; D019901:Locus Control Region / Q000235:genetics*; D051379:Mice; D054353:Perforin; D016133:Polymerase Chain Reaction; D052899:Pore Forming Cytotoxic Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
C510468:PRF1 protein, human; D052899:Pore Forming Cytotoxic Proteins; D054353:Perforin