Upstream elements bestow T-cell and haemopoietic progenitor-specific activity on the granzyme B promoter.
Cytotoxic T cells and early haemopoietic progenitors share the expression of a number of specific genes. Of these, granzyme B has attracted particular interest because of its role in inducing apoptosis during cytotoxic T cell-mediated target cell killing, and its potential role in the mobilisation and homeostasis of haemopoietic stem cells. Studies of granzyme B regulation should therefore yield valuable information concerning the molecular control of these processes, and also identify elements capable of directing gene expression to two cell types of relevance to gene therapy. Here we show that proximal regulatory elements already known to direct promoter activity in T cells are similarly active in haemopoietic progenitors. However, this activity is not strictly specific, since the promoter regions also direct low levels of reporter gene expression in fibroblasts. More importantly, we also report the presence of two previously unidentified clusters of DNaseI hypersensitive sites upstream from the murine granzyme B gene, and show that these regions impart both increased transcriptional activity and the appropriate cell type specificity on the granzyme B promoter. These upstream regulatory regions are therefore likely to play a key role in the coordination of granzyme B expression in vivo.
Section of Haemopoietic Cell and Gene Therapeutics, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester M20 4BX, UK.
B A
BA
Johnson
V A
VA
John
R
R
Henschler
I N
IN
Hampson
C M
CM
Heyworth
C K
CK
Babichuk
R C
RC
Bleackley
T M
TM
Dexter
M A
MA
Cross
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D003360:Cosmids; D003850:Deoxyribonuclease I / Q000378:metabolism; D053804:Granzymes; D006412:Hematopoietic Stem Cells / Q000276:immunology*; D008297:Male; D051379:Mice; D011401:Promoter Regions, Genetic; D012697:Serine Endopeptidases / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology*
D003850:Deoxyribonuclease I; D053804:Granzymes; C509214:Gzmb protein, mouse; D012697:Serine Endopeptidases