A critical role for C/EBPbeta binding to the AABS promoter response element in the human iNOS gene.

Zhong Guo, Lifang Shao, Xuesheng Feng, Kaye Reid, Eric Marderstein, Atsunori Nakao, David A. Geller

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

The human iNOS (hiNOS) gene is expressed in a tissue-specific manner, but the molecular basis for this regulation has not been elucidated. Here, we show that liver cell-specific hiNOS gene activation involves protein-DNA binding to an A-activator binding site (AABS) located at -192 nucleotides in the hiNOS promoter region. Mutation of this site in the -7.2 kb hiNOS promoter construct inhibited basal hiNOS promoter activity in primary rat hepatocytes (77%), and two human liver cell lines, AKN-1 (63%) and HepG2 (60%), but had no significant effect on basal hiNOS activity in three non-hepatic human cell types. Interestingly, mutation of AABS significantly abrogated cytokine-induced promoter activity in all cell types. C/EBPbeta transcription factor bound to AABS by gel shift assay. Overexpression of C/EBPbeta active form (LAP) increased hiNOS basal promoter activity approximately sixfold in liver cells, but had minimal effect in non-hepatic cells. In contrast, overexpression of the transcriptional inhibitor (LIP) strongly suppressed both basal and cytokine-inducible promoter activity. These data show that the cis-acting AABS DNA element mediates liver-specific basal hiNOS promoter activity through binding of the trans-acting C/EBPbeta factor. Further, C/EBPbeta binding to AABS functions as a "switchpoint" that is necessary for cytokine-inducible hiNOS gene expression in all cell types examined.

Original languageEnglish
Pages (from-to)1718-1720
Number of pages3
JournalFASEB Journal
Volume17
Issue number12
Publication statusPublished - 2003
Externally publishedYes

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CCAAT-Enhancer-Binding Protein-beta
Response Elements
Genes
Liver
Binding Sites
Cytokines
Cells
DNA-Binding Proteins
Genetic Promoter Regions
Gene expression
Rats
Assays
Transcription Factors
Nucleotides
Gels
Chemical activation
Tissue
Mutation
DNA
Human Activities

Cite this

Guo, Z., Shao, L., Feng, X., Reid, K., Marderstein, E., Nakao, A., & Geller, D. A. (2003). A critical role for C/EBPbeta binding to the AABS promoter response element in the human iNOS gene. FASEB Journal, 17(12), 1718-1720.

A critical role for C/EBPbeta binding to the AABS promoter response element in the human iNOS gene. / Guo, Zhong; Shao, Lifang; Feng, Xuesheng; Reid, Kaye; Marderstein, Eric; Nakao, Atsunori; Geller, David A.

In: FASEB Journal, Vol. 17, No. 12, 2003, p. 1718-1720.

Research output: Contribution to journalArticle

Guo, Z, Shao, L, Feng, X, Reid, K, Marderstein, E, Nakao, A & Geller, DA 2003, 'A critical role for C/EBPbeta binding to the AABS promoter response element in the human iNOS gene.', FASEB Journal, vol. 17, no. 12, pp. 1718-1720.
Guo, Zhong ; Shao, Lifang ; Feng, Xuesheng ; Reid, Kaye ; Marderstein, Eric ; Nakao, Atsunori ; Geller, David A. / A critical role for C/EBPbeta binding to the AABS promoter response element in the human iNOS gene. In: FASEB Journal. 2003 ; Vol. 17, No. 12. pp. 1718-1720.
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