A novel lineage-specific hypersensitive site is essential for position independent granzyme B expression in transgenic mice.
The granzyme B gene is activated upon cytotoxic T cell stimulation and the protein is a key inducer of apoptosis in target cells. Previous studies have identified important proximal regulatory regions but these proved insufficient to drive expression in vivo. We identified a DNase1 hypersensitive site (HS2) 3.9kb upstream of the transcription start site that was present in stimulated but not resting CD8+ cells. The CTL line CTLL R8 was stably transfected with GFP reporter constructs and showed consistently higher fluorescence values when HS2 was included. In transgenic mice the presence of the relevant region of DNA resulted in inducible, CTL-specific transcription of the transgene in all transgenic founder lines analyzed. Deletion of HS2 resulted in a 10-fold reduction in expression. This is the first report of a major distal regulatory element in the control of granzyme B transcription.
Duggan
Brenda L
BL
Department of Biochemistry, University of Alberta, Edmonton, Alta., Canada T6G 2H7.
Cabilio
Nora R
NR
Dickie
Peter
P
Witmer
Jennifer
J
Goping
Ing Swie
IS
Underhill
D Alan
DA
Bleackley
R Chris
RC
D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000502:physiology*; D002478:Cells, Cultured; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D053804:Granzymes / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D015533:Transcriptional Activation / Q000235:genetics*
D053804:Granzymes