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Vol. 12, No. 12, pp. 1787-1800, June 15, 1998
Cell Biology Program and
2 Molecular Biology Program, Nuclear receptors transduce hormonal signals by binding directly
to DNA target sites in promoters and modulating the transcription of
linked genes. Receptor-mediated transactivation appears to be
potentiated in response to ligand by a number of coactivators that may
provide key interactions with components of the transcription preinitiation complex and/or alter chromatin structure.
Here, we use the vitamin D3 receptor ligand-binding domain
(VDR LBD) as an affinity matrix to identify components of a
transcriptionally active nuclear extract that interact with VDR in
response to ligand. We describe the purification of a complex of at
least 10 VDR interacting proteins (DRIPs) ranging from 65 to 250 kD
that associate with the receptor in a strictly 1,25-dihydroxyvitamin
D3-dependent manner. These proteins also appear to interact
with other, but not all, nuclear receptors, such as the thyroid hormone
receptor. The DRIPs are distinct from known nuclear receptor
coactivators, although like these coactivators, their interaction also
requires the AF-2 transactivation motif of VDR. In addition, the DRIP
complex contains histone acetyltransferase activity, indicating that at least one or more of the DRIPs may function at the level of nucleosomal modification. However, we show that the DRIPs selectively enhance the
transcriptional activity of VDR on a naked DNA template utilizing a
cell-free, ligand-dependent transcription assay. Moreover, this activity can be specifically depleted from the extract by liganded, but
not unliganded, VDR-LBD. Overexpression of DRIP100 in vivo resulted in
a strong squelching of VDR transactivation, suggesting the
sequestration of other limiting factors, including components of the
DRIP complex. These results demonstrate the existence of a new complex
of novel functional nuclear receptor coactivators.
[Key Words:
Vitamin D3 receptor; ligand-binding domain; nuclear receptor coactivators; VDR
transactivation; cell-free transcription]
GENES & DEVELOPMENT 12:1787-1800 © 1998 by Cold Spring Harbor Laboratory Press ISSN 0890-9369/98 $5.00
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