Identification of a cell-specific transcription activation domain within the human Ah receptor nuclear translocator.
In the presence of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related chemicals, the Ah receptor nuclear translocator (Arnt) forms a heterodimeric complex with the ligand-bound Ah receptor, leading to recognition of dioxin-responsive elements within the enhancer of the CYP1A1 gene and transcription activation by an unknown mechanism. To understand the role of Arnt in transcription activation by the Ah receptor-Arnt heterodimer, we performed a deletion analysis of Arnt to locate domains that are directly involved in transcription activation. We showed that the C-terminal 34 amino acids of Arnt encode a transcription activation domain (TAD) that functions independently of other sequences in the Ah receptor complex when attached to the heterologous Gal4 DNA binding domain. Deletion of the C-terminal acidic-rich 14 amino acids completely abolishes activity. Sequences important in Arnt TAD function were independent of the glutamine-rich region which is an important structural feature in the TAD of other transcription factors. The strength of the Arnt TAD when compared with the strong TAD from the herpes simplex virus VP16 protein was cell-type specific. Both the Arnt and VP16 TAD were equally strong in COS-1 cells, but the Arnt TAD had weak activity in an Arnt-deficient mouse hepatoma cell line and was not needed for restoration of CYP1A1 activation. These results imply that for CYP1A1 activation the Ah receptor provides the dominant activation function for the heterodimer in hepatoma cells. The potential of the Arnt TAD to contribute to activation by the Ah receptor complex is likely determined by availability or activity of cell-specific factors with which the TAD interacts.
Corton
J C
JC
Chemical Industry Institute of Toxicology, Research Triangle Park, North Carolina 27709, USA.
Moreno
E S
ES
Hovis
S M
SM
Leonard
L S
LS
Gaido
K W
KW
Joyce
M M
MM
Kennett
S B
SB
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D051784:Aryl Hydrocarbon Receptor Nuclear Translocator; D001483:Base Sequence; D051792:Basic Helix-Loop-Helix Proteins; D002478:Cells, Cultured; D002874:Chromosome Mapping; D006801:Humans; D008114:Liver Neoplasms, Experimental / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D018336:Receptors, Aryl Hydrocarbon / Q000737:chemistry / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D015534:Trans-Activators / Q000737:chemistry*; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics*; D015533:Transcriptional Activation / Q000235:genetics*
D000596:Amino Acids; D051784:Aryl Hydrocarbon Receptor Nuclear Translocator; D051792:Basic Helix-Loop-Helix Proteins; D018336:Receptors, Aryl Hydrocarbon; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C490989:ARNT2 protein, human; C099817:Arnt2 protein, mouse