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(Journal of Leukocyte Biology. 2002;71:890-904.)
© 2002 by Society for Leukocyte Biology

Regulation of NRAMP1 gene expression by 1,25-dihydroxy-vitamin D₃ in HL-60 phagocytes

E. A. Roig^*, E. Richer^*, F. Canonne-Hergaux, P. Gros and M. F. M. Cellier^*

^* INRS-Institut Armand-Frappier, Laval, PQ, Canada; and
Department of Biochemistry, McGill University, Montréal, PQ, Canada

Correspondence: M. F. M. Cellier, INRS-Institut Armand-Frappier, 531 Bd des prairies, Laval, H7V 1B7, PQ, Canada. E-mail: mathieu.cellier{at}inrs-iaf.uquebec.ca

The natural resistance-associated macrophage protein 1 (Nramp1) is a proton-dependent transporter of divalent metals. We studied NRAMP1 expression during HL-60 differentiation induced by VD and VD agonists. NRAMP1 and CD14 gene expression differed in kinetics of induction, mRNA levels and stability, and response to VD combined with PMA, whereas a combination of VD and IFN- induced similar up-regulation. NRAMP1 protein expression paralleled the accumulation of mRNA and was localized in the phagosomal membrane after phagocytosis. A promoter construct extending 647 bp upstream of NRAMP1 ATG showed myeloid-specific transcription in transient transfection assays, which was up-regulated by VD in HL-60. In HL-60 clones stably transfected with this construct, transcription was apparently induced through indirect VD genomic effects, and there was accordance between the levels of reporter transcription and endogenous NRAMP1 mRNA in response to VD but not to IFN-. Thus, VD genomic effects stimulate NRAMP1 transcription and protein expression in maturing phagocytes.

Key Words: monocytic differentiation • interferon- • Northern blot • immunofluorescence • luciferase reporter gene

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