Cloning and characterization of granzyme B gene promoter

Conference · Fri Mar 15 04:00:00 EST 1991 · FASEB Journal (Federation of American Societies for Experimental Biology); (United States)

OSTI ID:5239527

Jiang, M; Huff, M; Showe, L; Weinmann, R; Trinchieri, G ^[1]; McMahon, S ^[2]

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1. Wistar Inst., Philadelphia, PA (United States)
2. Univ. of Pennsylvania, Philadelphia (United States)

Cytolytic T cells and NK cells contain specific serine esterases (Granzymes) that are not known to be expressed by other cell types. The human genes coding for two of these esterases, granzyme A and granzyme B, have been cloned. In vitro activation by mitogens, phorbol diesters, IL-2 and TCR/CD3 ligands induces increased accumulation of granzyme mRNA in T cells. The authors have isolated human genomic sequences encoding the granzyme B gene from a {lambda}fix DNA library. They have localized the transcription initiation site using primer extension. The 5{prime} flanking region of this gene was subcloned into a vector containing the luciferase reporter coding sequences. This construct was transfected by electroporation into human peripheral blood lymphocytes (PBL) pretreated for 48 h with PHA. A low level of luciferase activity was detected in cell extracts of transfected cells. Luciferase activity was greatly enhanced by stimulation of the transfected cells with PHA, TPA, and the anti-CD3 antibody OKT3, but not by stimulation with IL-2. Thus, this 5{prime} upstream sequence contains a functional granzyme B gene promoter which responds to several of the same stimuli which result in increased level of granzyme B mRNA from the endogenous gene.

• OSTI ID: 5239527
• Report Number(s): CONF-9104107--
• Conference Information: Journal Name: FASEB Journal (Federation of American Societies for Experimental Biology); (United States) Journal Volume: 5:5
• Country of Publication: United States
• Language: English

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63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AMINO ACID SEQUENCE
ANIMAL CELLS
ANIMALS
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL EFFECTS
BIOLOGICAL MATERIALS
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ESTERS
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LEUKOCYTES
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LYMPHOCYTES
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MAN
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MESSENGER-RNA
MITOGENS
MOLECULAR STRUCTURE
NUCLEIC ACIDS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
PHORBOL ESTERS
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