The effect of the synthetic glucocorticoid hormone dexamethasone on human gamma chain fibrinogen gene expression was examined. The whole promoter region of 3.8 kb of this gene and progressive 5'-deletions were inserted into a promoterless expression vector, upstream of the luciferase gene and transiently transfected into the human hepatoma HepG2 cells, in the presence or in the absence of dexamethasone stimulation. Deletion analysis allowed to identify a region located between -1359 and -954 bp upstream from the transcription start site, involved in hormone inducibility. On the basis of a computer-assisted analysis, a putative GRE was found in this region at bases -1116 to -1102. Specific point mutations eliminating this putative GRE led to complete loss of glucocorticoid inducibility, thus indicating its functional role. Binding of the rat glucocorticoid receptor to this site was demonstrated by mobility-shift assays.

Identification of a glucocorticoid response element in the human gamma chain fibrinogen promoter / R. Asselta, S. Duga, M. Modugno, M. Malcovati, M.L. Tenchini. - In: THROMBOSIS AND HAEMOSTASIS. - ISSN 0340-6245. - 79:6(1998 Jun), pp. 1144-1150.

Identification of a glucocorticoid response element in the human gamma chain fibrinogen promoter

R. Asselta
Primo
;
S. Duga
Secondo
;
1998

Abstract

The effect of the synthetic glucocorticoid hormone dexamethasone on human gamma chain fibrinogen gene expression was examined. The whole promoter region of 3.8 kb of this gene and progressive 5'-deletions were inserted into a promoterless expression vector, upstream of the luciferase gene and transiently transfected into the human hepatoma HepG2 cells, in the presence or in the absence of dexamethasone stimulation. Deletion analysis allowed to identify a region located between -1359 and -954 bp upstream from the transcription start site, involved in hormone inducibility. On the basis of a computer-assisted analysis, a putative GRE was found in this region at bases -1116 to -1102. Specific point mutations eliminating this putative GRE led to complete loss of glucocorticoid inducibility, thus indicating its functional role. Binding of the rat glucocorticoid receptor to this site was demonstrated by mobility-shift assays.
Dexamethasone ; Animals ; Carcinoma, Hepatocellular ; DNA Mutational Analysis ; DNA-Binding Proteins ; Humans ; Fibrinogen ; Receptors, Glucocorticoid ; Recombinant Fusion Proteins ; Cloning, Molecular ; Rats ; Mutagenesis, Site-Directed ; Liver Neoplasms ; Promoter Regions, Genetic ; Tumor Cells, Cultured ; Genetic Vectors ; Gene Expression Regulation ; Consensus Sequence ; Gene Library
Settore BIO/11 - Biologia Molecolare
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/209586
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