Identification of human granzyme B promoter regulatory elements interacting with activated T-cell-specific proteins: implication of Ikaros and CBF binding sites in promoter activation.

Abstract

Granzyme B serine protease is found in the granules of activated cytotoxic T cells and in natural and lymphokine-activated killer cells. This protease plays a critical role in the rapid induction of target cell DNA fragmentation. The DNA regulatory elements that are responsible for the specificity of granzyme B gene transcription in activated T-cells reside between nt -148 and +60 (relative to the transcription start point at +1) of the human granzyme B gene promoter. This region contains binding sites for the transcription factors Ikaros, CBF, Ets, and AP-1. Mutational analysis of the human granzyme B promoter reveals that the Ikaros binding site (-143 to -114) and the AP-1/CBF binding site (-103 to -77) are essential for the activation of transcription in phytohemagglutinin-activated peripheral blood lymphocytes, whereas mutation of the Ets binding site does not affect promoter activity in these cells.

Death by a Thousand Cuts: Granzyme Pathways of Programmed Cell Death

“…Analysis of reporter assays using promoters that had been systematically mutated at these sites in primary cells and cell lines revealed subtle differences in the importance of some transcription factors in primary cells versus cell lines. For example, AP-1, CREB, and CBF were not as important for transcription in primary cells as they appeared to be in cell lines (37,39). These studies suggested that combinations of transcription factors (particularly AP-1 and CBF) are required to activate GzmB expression in primary cells.…”

“…The most compelling difference between the mouse and human GzmB gene promoter is the importance of the Ikaros site in human GzmB expression. Mutations introduced into the Ikarosbinding site of the human, but not mouse, GzmB promoter abrogate expression in primary T cells (37,39).…”

“…This region contains binding sites for two ubiquitous transcription factors, activating transcription factor/cyclic AMP-responsive element binding protein (ATF/CREB) and activator protein-1 (AP-1), and two lymphoidspecific factors, Ikaros and core-binding factor (CBF/PEBP2) (36). Several of these transcription factor-binding sites are evolutionarily conserved between the human and mouse GzmB promoters (37,38). Analysis of reporter assays using promoters that had been systematically mutated at these sites in primary cells and cell lines revealed subtle differences in the importance of some transcription factors in primary cells versus cell lines.…”

Transcriptional regulation of osteopontin gene in vivo by PEBP2αA/CBFA1 and ETS1 in the skeletal tissues

“…This domain mediates DNA binding as well as dimerization to PEBP2b/CBFB, which has no direct DNA binding ability, and confers enhanced DNA binding capacity to PEBP2a (Ogawa et al, 1993b;Wang et al, 1993). PEBP2a speci®cally recognizes a consensus sequence, PuACCPuCA and has been reported to control polyoma virus enhancer (Kamachi et al, 1990), murine leukemia virus enhancers , T cell-speci®c genes (Giese et al, 1995;Hallberg et al, 1992;Hsiang et al, 1993;Prosser et al, 1992;Redondo et al, 1992), enzymes (myeloperoxidase, neutrophil elastase, granzyme B serine protease) (Nuchprayoon et al, 1994;Wargnier et al, 1995) and cytokines and their receptors (GM-CSF, IL3, CSF-1) (Cameron et al, 1994;Frank et al, 1995;Zhang et al, 1994). PEBP2aA and ETS1 have been shown to bind together and interact synergistically on the promoter of the T cell receptor a gene (Giese et al, 1995) and T cell receptor b gene (Wotton et al, 1994).…”

UVA Induces Granzyme B in Human Keratinocytes through MIF

“…First, MIF and UVA were found to activate ERK, p38 MAPK, and JNK modules (13,14). Second, MAPK are candidates for the regulation of AP1 and CREB, two regulatory elements of GrB promoter (15,16). Third, we have already demonstrated that UVB induced GrB expression via MAPK pathway in keratinocytes (6).…”