TY - JOUR
T1 - Association of prokaryotic and eukaryotic chaperone proteins with the human 1α,25-dihydroxyvitamin D3 receptor
AU - Craig, Theodore A.
AU - Lutz, Ward H.
AU - Kumar, Rajiv
N1 - Funding Information:
We thank Dr. David Toft for useful discussions. This study was supported by NIH DK25409.
PY - 1999/7/5
Y1 - 1999/7/5
N2 - Steroid hormone receptors (SHR) form complexes with heat shock proteins (hsps). The 1α,25-dihydroxyvitamin D3 receptor (VDR) has not been previously shown to interact with hsps. During expression and purification of VDR-glutathione S-transferase (VDR-GST) fusion proteins encompassing full-length, DNA, and ligand-binding domains of the VDR (FL-VDR, DBD-VDR, and LBD-VDR), we observed binding of bacterial hsps with VDR-GST constructs. All VDR constructs bound DnaK in amounts greater than GST alone and bound smaller amounts of DnaJ or GrpE. GroEL bound only to FL-VDR. GroES did not bind to VDR. When VDR-GST constructs were incubated with a reticulocyte lysate system that has been used previously to examine SHR-hsp interactions, eukaryotic hsc70 was detected bound to FL-VDR and DBD-VDR. Binding of hsp90 to VDR was not detected. However, geldanamycin, an hsp90 inhibitor, reduced 1α,25-dihydroxyvitamin D3-mediated gene activation in osteoblasts. Our data show that the bacterial and eukaryotic hsps associate with the VDR and might be involved in VDR function.
AB - Steroid hormone receptors (SHR) form complexes with heat shock proteins (hsps). The 1α,25-dihydroxyvitamin D3 receptor (VDR) has not been previously shown to interact with hsps. During expression and purification of VDR-glutathione S-transferase (VDR-GST) fusion proteins encompassing full-length, DNA, and ligand-binding domains of the VDR (FL-VDR, DBD-VDR, and LBD-VDR), we observed binding of bacterial hsps with VDR-GST constructs. All VDR constructs bound DnaK in amounts greater than GST alone and bound smaller amounts of DnaJ or GrpE. GroEL bound only to FL-VDR. GroES did not bind to VDR. When VDR-GST constructs were incubated with a reticulocyte lysate system that has been used previously to examine SHR-hsp interactions, eukaryotic hsc70 was detected bound to FL-VDR and DBD-VDR. Binding of hsp90 to VDR was not detected. However, geldanamycin, an hsp90 inhibitor, reduced 1α,25-dihydroxyvitamin D3-mediated gene activation in osteoblasts. Our data show that the bacterial and eukaryotic hsps associate with the VDR and might be involved in VDR function.
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U2 - 10.1006/bbrc.1999.0931
DO - 10.1006/bbrc.1999.0931
M3 - Article
C2 - 10403788
AN - SCOPUS:0033526834
SN - 0006-291X
VL - 260
SP - 446
EP - 452
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 2
ER -