TY - JOUR
T1 - Regulation of avian calbindin-d28k gene expression in primary chick kidney cells
T2 - Importance of posttranscriptional mechanisms and calcium ion concentration
AU - Enomoto, Hiroshi
AU - Hendy, Geoffrey N.
AU - Andrew, Glen K.
AU - Clemens, Thomas L.
N1 - Copyright:
Copyright 2016 Elsevier B.V., All rights reserved.
PY - 1992/6
Y1 - 1992/6
N2 - Vitamin D-dependent calcium-binding protein (calbindin-D28K). is regulated bv 1.25~dihydroxyvitamin Da [1, 25-(OH)2D3], andseveraiother factors in a-tissue-specific manner, but the controlling mechanisms are still poorly understood. In this study we examined the relative contributions of transcriptional and posttranscriptional mechanisms in the 1, 25-(OH)2D, control of calbindin-DZ8k mRNA expression in primary chick kidney cells and investigated the effect of extracellular Ca2+ on calbindin-D2sk gene expression in the presence and absence of hormone. 1, 25-(OH2)D3 treatment (lo-@ M) of cells grown in serum-free medium resulted in a marked 20-to 30-fold increase in calbindin-D28K mRNA peaking at 12-18 h, which then rapidly declined to basal levels by 24 h. The abrupt decline in mRNA appeared to be associated with a reduction in size of the calbindin-D28K transcripts. Nuclear run-off assays showed only a slight (I.5-fold) increase in calbindin-D28K gene transcription 2 h after 1, 25-(OH2)D3, whereas parallel assays clearly demonstrated a marked (7-fold) induction in the rate of metallothionein gene transcription 2 h after treatment of chick kidney cells with 10 pM zinc. The induction of calbindin-D mRNA by 1, 25-(OH)2D3 required ongoing protein synthesis, since it was blocked by cycloheximide. Calbindin-Dssk mRNA was stable for 12 h in the presence of actinomycin-D in both vitamin D-deficient and 1, 25-(OH)2D3-treated cells. Both basal and 1, 25-(OH)2D3-induced calbindin-D28K mRNA were modulated by the extracellular Ca2+, with maximum expression occurring at l-2 mM. We conclude that 1, 25-(OH)2D3 induces kidney calbindin-D, s mRNA by producing a small increase in its transcriptional rate, which is accompanied by pronounced posttranscriptional effects(s). The striking modulation of calbindin-D28K expression by extracellular Ca2+ is consistent with a putative role for this protein in the regulation of this ion in the kidney cell.
AB - Vitamin D-dependent calcium-binding protein (calbindin-D28K). is regulated bv 1.25~dihydroxyvitamin Da [1, 25-(OH)2D3], andseveraiother factors in a-tissue-specific manner, but the controlling mechanisms are still poorly understood. In this study we examined the relative contributions of transcriptional and posttranscriptional mechanisms in the 1, 25-(OH)2D, control of calbindin-DZ8k mRNA expression in primary chick kidney cells and investigated the effect of extracellular Ca2+ on calbindin-D2sk gene expression in the presence and absence of hormone. 1, 25-(OH2)D3 treatment (lo-@ M) of cells grown in serum-free medium resulted in a marked 20-to 30-fold increase in calbindin-D28K mRNA peaking at 12-18 h, which then rapidly declined to basal levels by 24 h. The abrupt decline in mRNA appeared to be associated with a reduction in size of the calbindin-D28K transcripts. Nuclear run-off assays showed only a slight (I.5-fold) increase in calbindin-D28K gene transcription 2 h after 1, 25-(OH2)D3, whereas parallel assays clearly demonstrated a marked (7-fold) induction in the rate of metallothionein gene transcription 2 h after treatment of chick kidney cells with 10 pM zinc. The induction of calbindin-D mRNA by 1, 25-(OH)2D3 required ongoing protein synthesis, since it was blocked by cycloheximide. Calbindin-Dssk mRNA was stable for 12 h in the presence of actinomycin-D in both vitamin D-deficient and 1, 25-(OH)2D3-treated cells. Both basal and 1, 25-(OH)2D3-induced calbindin-D28K mRNA were modulated by the extracellular Ca2+, with maximum expression occurring at l-2 mM. We conclude that 1, 25-(OH)2D3 induces kidney calbindin-D, s mRNA by producing a small increase in its transcriptional rate, which is accompanied by pronounced posttranscriptional effects(s). The striking modulation of calbindin-D28K expression by extracellular Ca2+ is consistent with a putative role for this protein in the regulation of this ion in the kidney cell.
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U2 - 10.1210/endo.130.6.1375904
DO - 10.1210/endo.130.6.1375904
M3 - Article
C2 - 1375904
AN - SCOPUS:0026692189
SN - 0013-7227
VL - 130
SP - 3467
EP - 3474
JO - Endocrinology
JF - Endocrinology
IS - 6
ER -