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(Received for publication, March 6, 1996, and in revised form, April 15, 1996)
From the Department of Biochemistry, University of Alberta,
Edmonton, Alberta T6G 2H7, Canada
A murine granzyme B promoter fragment that
extends 243 base pairs upstream of the transcription start site confers
high levels of luciferase reporter gene activity in transient
transfection assays into T cells and mouse L cell fibroblasts. This
promoter fragment contains canonical binding sites for the
transcription factors AP-1, core binding factor (CBF), Ikaros, and the
cyclic AMP responsive element binding protein (CREB). Oligonucleotides
containing the granzyme B AP-1 or CBF elements form specific complexes
with proteins present in nuclear extracts from activated
CD8+ splenocytes, MTL cells, EL4 T cells, and L cells. A
strong DNase1 hypersensitive site that coincides with the closely
associated AP-1, CBF, Ikaros, and CRE elements is present in activated
CD8+ T cells but not in resting T cells or L cells. Both
in vitro and in vivo footprints are observed at
these sequence elements in activated cytotoxic T cells (CTL) but not in
resting T cells. The endogenous granzyme B gene is CTL-specific as no
mRNA is detectable in EL4 or L cells. We propose that a condensed
chromatin structure at the granzyme B promoter is responsible for
transcription factor inaccessibility and repression of transcription in
non-T cells.
Volume 271, Number 28,
Issue of July 12, 1996
pp. 16485-16493
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
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