Regulation of avian calbindin-d28k gene expression in primary chick kidney cells: Importance of posttranscriptional mechanisms and calcium ion concentration

Hiroshi Enomoto, Geoffrey N. Hendy, Glen K. Andrew, Thomas L. Clemens

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)3467-3474
Number of pages8
JournalEndocrinology
Volume130
Issue number6
DOIs
StatePublished - Jun 1992
Externally publishedYes

ASJC Scopus subject areas

  • Endocrinology

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