The chicken malic enzyme gene: Structural organization and identification of triiodothyronine response elements in the 5'-flanking DNA

Dean W. Hodnett, Dominic A. Fantozzi, Debbie C. Thurmond, Stephen A. Klautky, Kathleen G. MacPhee, Shawn T. Estrem, Gang Xu, Alan G. Goodridge

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

In vivo, feeding stimulates and starvation inhibits transcription of the malic enzyme gene. In chick-embryo hepatocytes in culture, triiodothyronine (T3) stimulates and glucagon inhibits transcription of this gene. As a first step in the characterization of the involved regulatory mechanisms, fragments of genomic DNA spanning the structural and 5'-flanking regions of the chicken malic enzyme gene were cloned. The coding region of the gene is organized into 14 exons and 13 introns and is greater than 106 kb in length. The size of the gene, the number and lengths of the exons, and positions at which introns are inserted into the coding regions are virtually identical in the chicken and rat genes. When transiently transfected into chick-embryo hepatocytes, 5800 bp of 5'-flanking DNA conferred T3 responsiveness to a linked chloramphenicol acetyltransferase (CAT) reporter gene. Using deletion and site-specific mutations of 5'-flanking DNA, we identified a complex T3 response unit that contains one major T3 response element (T3RE) and several minor ones. The major element contains two degenerate copies of the hexamer, RGGWMA, separated by 4 bp and was a strong repressor in the absence of ligand. Endogenous levels of T3 receptor are sufficient to allow the T3 response elements in the upstream region of the malic enzyme gene to confer responsiveness to T3, suggesting that they are physiologically relevant.

Original languageEnglish (US)
Pages (from-to)309-324
Number of pages16
JournalArchives of Biochemistry and Biophysics
Volume334
Issue number2
DOIs
StatePublished - Oct 15 1996
Externally publishedYes

Fingerprint

Response Elements
Triiodothyronine
Chickens
Genes
DNA
Enzymes
Chick Embryo
Introns
Transcription
Hepatocytes
Exons
CD3 Antigens
Thyroid Hormone Receptors
Chloramphenicol O-Acetyltransferase
5' Flanking Region
Starvation
Glucagon
Reporter Genes
Cell culture
Rats

Keywords

  • avian
  • malic enzyme gene
  • promoter
  • thyroid hormone
  • transcription

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Hodnett, D. W., Fantozzi, D. A., Thurmond, D. C., Klautky, S. A., MacPhee, K. G., Estrem, S. T., ... Goodridge, A. G. (1996). The chicken malic enzyme gene: Structural organization and identification of triiodothyronine response elements in the 5'-flanking DNA. Archives of Biochemistry and Biophysics, 334(2), 309-324. https://doi.org/10.1006/abbi.1996.0460

The chicken malic enzyme gene : Structural organization and identification of triiodothyronine response elements in the 5'-flanking DNA. / Hodnett, Dean W.; Fantozzi, Dominic A.; Thurmond, Debbie C.; Klautky, Stephen A.; MacPhee, Kathleen G.; Estrem, Shawn T.; Xu, Gang; Goodridge, Alan G.

In: Archives of Biochemistry and Biophysics, Vol. 334, No. 2, 15.10.1996, p. 309-324.

Research output: Contribution to journalArticle

Hodnett, DW, Fantozzi, DA, Thurmond, DC, Klautky, SA, MacPhee, KG, Estrem, ST, Xu, G & Goodridge, AG 1996, 'The chicken malic enzyme gene: Structural organization and identification of triiodothyronine response elements in the 5'-flanking DNA', Archives of Biochemistry and Biophysics, vol. 334, no. 2, pp. 309-324. https://doi.org/10.1006/abbi.1996.0460
Hodnett, Dean W. ; Fantozzi, Dominic A. ; Thurmond, Debbie C. ; Klautky, Stephen A. ; MacPhee, Kathleen G. ; Estrem, Shawn T. ; Xu, Gang ; Goodridge, Alan G. / The chicken malic enzyme gene : Structural organization and identification of triiodothyronine response elements in the 5'-flanking DNA. In: Archives of Biochemistry and Biophysics. 1996 ; Vol. 334, No. 2. pp. 309-324.
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