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CCAAT/enhancer binding protein beta mediates the activation of the murine alpha1(I) collagen promoter by acetaldehyde.
Arch Biochem Biophys. 2000 Jun 01; 378(1):57-64.AB

Abstract

Acetaldehyde was previously shown to activate the alpha1(I) and alpha2(I) collagen promoters and to increase collagen production in activated stellate cells. Also, CCAAT/enhancer binding protein beta (C/EBPbeta) binds and activates the mouse alpha1(I) collagen promoter. This study investigates the role of C/EBPbeta in mediating the activation of the alpha1(I) collagen promoter by acetaldehyde. Nuclear extracts isolated from cultured activated rat hepatic stellate cells formed four protein-DNA complexes on electrophoretic mobility shift assay with an oligonucleotide including the C/EBP binding site between -365 and -335 in the alpha1(I) collagen promoter. The four complexes were identified to represent C/EBPbeta binding to the oligonucleotide by supershift with C/EBPbeta antibody. The principal C/EBP isoform found in the nuclear extracts from stellate cells was C/EBPbeta, with very low amounts of C/EBPalpha detected. Acetaldehyde (200 microM) increased C/EBPbeta protein in stellate nuclear extracts, increased its binding to the promoter, and activated the alpha1(I) collagen promoter in transfected stellate cells. Mutation of the C/EBPbeta binding site markedly decreased nuclear protein binding. A transfected promoter, mutated at the C/EBP binding site, had decreased basal activity, was not activated by acetaldehyde, and was not activated when cotransfected with a C/EBPbeta expression vector. This study shows that C/EBPbeta is the predominant C/EBP isoform found in activated stellate cells and that increased C/EBPbeta protein and C/EBPbeta binding to a proximal C/EBP binding site in the promoter mediates the activating effect of acetaldehyde.

Authors+Show Affiliations

Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

10871045

Citation

Attard, F A., et al. "CCAAT/enhancer Binding Protein Beta Mediates the Activation of the Murine alpha1(I) Collagen Promoter By Acetaldehyde." Archives of Biochemistry and Biophysics, vol. 378, no. 1, 2000, pp. 57-64.
Attard FA, Wang L, Potter JJ, et al. CCAAT/enhancer binding protein beta mediates the activation of the murine alpha1(I) collagen promoter by acetaldehyde. Arch Biochem Biophys. 2000;378(1):57-64.
Attard, F. A., Wang, L., Potter, J. J., Rennie-Tankersley, L., & Mezey, E. (2000). CCAAT/enhancer binding protein beta mediates the activation of the murine alpha1(I) collagen promoter by acetaldehyde. Archives of Biochemistry and Biophysics, 378(1), 57-64.
Attard FA, et al. CCAAT/enhancer Binding Protein Beta Mediates the Activation of the Murine alpha1(I) Collagen Promoter By Acetaldehyde. Arch Biochem Biophys. 2000 Jun 1;378(1):57-64. PubMed PMID: 10871045.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - CCAAT/enhancer binding protein beta mediates the activation of the murine alpha1(I) collagen promoter by acetaldehyde. AU - Attard,F A, AU - Wang,L, AU - Potter,J J, AU - Rennie-Tankersley,L, AU - Mezey,E, PY - 2000/6/28/pubmed PY - 2000/7/25/medline PY - 2000/6/28/entrez SP - 57 EP - 64 JF - Archives of biochemistry and biophysics JO - Arch Biochem Biophys VL - 378 IS - 1 N2 - Acetaldehyde was previously shown to activate the alpha1(I) and alpha2(I) collagen promoters and to increase collagen production in activated stellate cells. Also, CCAAT/enhancer binding protein beta (C/EBPbeta) binds and activates the mouse alpha1(I) collagen promoter. This study investigates the role of C/EBPbeta in mediating the activation of the alpha1(I) collagen promoter by acetaldehyde. Nuclear extracts isolated from cultured activated rat hepatic stellate cells formed four protein-DNA complexes on electrophoretic mobility shift assay with an oligonucleotide including the C/EBP binding site between -365 and -335 in the alpha1(I) collagen promoter. The four complexes were identified to represent C/EBPbeta binding to the oligonucleotide by supershift with C/EBPbeta antibody. The principal C/EBP isoform found in the nuclear extracts from stellate cells was C/EBPbeta, with very low amounts of C/EBPalpha detected. Acetaldehyde (200 microM) increased C/EBPbeta protein in stellate nuclear extracts, increased its binding to the promoter, and activated the alpha1(I) collagen promoter in transfected stellate cells. Mutation of the C/EBPbeta binding site markedly decreased nuclear protein binding. A transfected promoter, mutated at the C/EBP binding site, had decreased basal activity, was not activated by acetaldehyde, and was not activated when cotransfected with a C/EBPbeta expression vector. This study shows that C/EBPbeta is the predominant C/EBP isoform found in activated stellate cells and that increased C/EBPbeta protein and C/EBPbeta binding to a proximal C/EBP binding site in the promoter mediates the activating effect of acetaldehyde. SN - 0003-9861 UR - https://www.unboundmedicine.com/medline/citation/10871045/CCAAT/enhancer_binding_protein_beta_mediates_the_activation_of_the_murine_alpha1_I__collagen_promoter_by_acetaldehyde_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-9861(00)91803-4 DB - PRIME DP - Unbound Medicine ER -