title: A novel lineage-specific hypersensitive site is essential for position independent granzyme B expression in transgenic mice.
abstract: 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.
authors.affiliation: Department of Biochemistry, University of Alberta, Edmonton, Alta., Canada T6G 2H7.
authors.forename: Brenda L
authors.initials: BL
authors.lastname: Duggan
authors.forename: Nora R
authors.initials: NR
authors.lastname: Cabilio
authors.forename: Peter
authors.initials: P
authors.lastname: Dickie
authors.forename: Jennifer
authors.initials: J
authors.lastname: Witmer
authors.forename: Ing Swie
authors.initials: IS
authors.lastname: Goping
authors.forename: D Alan
authors.initials: DA
authors.lastname: Underhill
authors.forename: R Chris
authors.initials: RC
authors.lastname: Bleackley
mesh_terms: 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*
chemical_list: D053804:Granzymes