Cloning, sequencing, and functional characterization of the vitamin D receptor in vitamin D-resistant New World primates

Abstract

New World primates (NWPs) have high circulating 1,25-dihydroxyvitamin D (1,25-(OH)2D) levels. Comparable levels would be harmful to Old World primates (OWPs) and humans. Thus, NWPs must have developed mechanisms of 1,25-(OH)2D resistance to survive. In humans, patients with hypocalcemic vitamin D-resistant rickets type II have high circulating vitamin D levels and vitamin D resistance due to expression of a dysfunctional vitamin D receptor (VDR). To examine if this could wholly or in part explain vitamin D resistance in NWPs, VDR from Saguinus oedipus (cotton top tamarin) NWP B95-8 cells was cloned by reverse-transcription polymerase chain reaction (RT-PCR). The NWP VDR cDNA sequence showed 96% homology at the DNA level and 98% homology at the amino acid level compared to human VDR. To assay for function, NWP VDR cDNA was transiently transfected into CV-1 cells with a vitamin D response element reporter plasmid. No difference between OWP and NWP VDR-directed transactivation was observed. These results indicate that the mechanism of vitamin D resistance in NWPs is not due to a dysfunctional VDR, and is consistent with our hypothesis that vitamin D resistance in NWPs is mediated by overexpression of a VDR-independent vitamin D response element binding protein.