The human transcription factor IID subunit human TATA-binding protein-associated factor 28 interacts in a ligand-reversible manner with the vitamin D(3) and thyroid hormone receptors.
Using coexpression in COS cells, we have identified novel interactions between the human TATA-binding protein-associated factor 28 (hTAF(II)28) component of transcription factor IID and the ligand binding domains (LBDs) of the nuclear receptors for vitamin D3 (VDR) and thyroid hormone (TRalpha). Interaction between hTAF(II)28 and the VDR and TR LBDs was ligand-reversible, whereas no interactions between hTAF(II)28 and the retinoid X receptors (RXRs) or other receptors were observed. TAF(II)28 interacted with two regions of the VDR, a 40-amino acid region spanning alpha-helices H3-H5 and alpha-helix H8. Interactions were also observed with the H3-H5 region of the TRalpha but not with the equivalent highly related region of the RXRgamma. Fine mapping using RXR derivatives in which single amino acids of the RXRgamma LBD have been replaced with their VDR counterparts shows that the determinants for interaction with hTAF(II)28 are located in alpha-helix H3 and are not identical to those previously identified for interactions with hTAF(II)55. We also describe a mutation in the H3-H5 region of the VDR LBD, which abolishes transactivation, and we show that interaction of hTAF(II)28 with this mutant is no longer ligand-reversible.
Mengus
G
G
Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS, INSERM, Université Louis Pastéur, BP-163-67404 Illkirch Cédex, France.
Gangloff
Y G
YG
Carré
L
L
Lavigne
A C
AC
Davidson
I
I
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D018167:Receptors, Calcitriol / Q000737:chemistry* / Q000378:metabolism*; D018168:Receptors, Retinoic Acid / Q000378:metabolism; D011988:Receptors, Thyroid Hormone / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D047488:Retinoid X Receptors; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D035381:TATA-Binding Protein Associated Factors; D035362:Transcription Factor TFIID; D014157:Transcription Factors / Q000378:metabolism*; D019473:Transcription Factors, TFII / Q000378:metabolism*; D014162:Transfection
D004268:DNA-Binding Proteins; D008024:Ligands; D018167:Receptors, Calcitriol; D018168:Receptors, Retinoic Acid; D011988:Receptors, Thyroid Hormone; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D047488:Retinoid X Receptors; C113944:TAF11 protein, human; D035381:TATA-Binding Protein Associated Factors; D035362:Transcription Factor TFIID; D014157:Transcription Factors; D019473:Transcription Factors, TFII