PMC8170129_01
The R80W mutation abolishes the transcriptional activity without affecting the structure or expression of VDRR80W
(A) Localization of the C238T mutation to exon 3 in the VDR gene resulting in the R80W in the C-terminal part of the second zinc finger in the DBD of the VDR. The amino acids surrounding the R80 in the VDR from different species with the R80W at the bottom. (B) Family tree where circles represent females, squares represent males, homozygous VDRR80W marked as filled black, heterozygous VDRWT/R80W marked as filled black and white, and homozygous VDRWT marked as filled white. (C, D)
In silico structure of the zinc fingers of the VDR interacting with DNA. (C) VDRWT with the R80 in red and with the hydrogen bonds between R80 and the DNA backbone indicated as blue lines. (D) The same region of VDRR80W with the W80 in blue. (E) VDR mRNA, (F) VDR protein and (G) CYP24A1 mRNA in VDRWT (black circles) and VDRR80W (white squares) CD4+ T cells activated in the presence of the indicated concentration of 25(OH)D3. (E-G) The expression levels are given as fold change normalized to the average expression level of VDRWT in control cells activated in the absence of 25(OH)D3. (E) Mean +- SD (VDRWT n = 6; VDRR80W n = 1). (F) Relative VDR protein expression as determined by the density of the VDR bands from Western blotting analysis of VDRWT and VDRR80W. The upper panel gives the density of VDRR80W (white squares) normalized to the average density of VDRWT (black circles) from control cells. The lower panel shows one representative Western blotting analysis out of five independent experiments of VDR and GAPDH (loading control) from three controls and the patient. Mean + SD (VDRWT n = 13; VDRR80W n = 1 repeated 5 times). (G) Mean +- SD (VDRWT n = 3; VDRR80W n = 1). n.s. means not significant.
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At the physical examination, the patient appeared normal except for alopecia and short stature with a height and weight below the 3% percentile ( Supplementary Figure 1 ). She often complained of bone pains in the arms and legs but otherwise she was doing well, and she partially catched up for her low stature, body weight and delayed bone age ( Supplementary Figure 1 ).