Journal of Leukocyte Biology Myeloid cells, immune suppression, tumor immunology
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(Journal of Leukocyte Biology. 2001;70:812-820.)
© 2001 by Society for Leukocyte Biology

A highly conserved c-fms gene intronic element controls macrophage-specific and regulated expression

S. Roy Himes*, Hiromi Tagoh{dagger}, Nilukshi Goonetilleke*, Tedjo Sasmono*, Delvac Oceandy*, Richard Clark*, Constanze Bonifer{dagger} and David A. Hume*

* Institute for Molecular Bioscience, University of Queensland, Brisbane, Australia, 4072 and
{dagger} University of Leeds, Molecular Medicine Unit, St. James University Hospital, Leeds, United Kingdom

Correspondence: Professor David A. Hume, Institute for Molecular Bioscience, University of Queensland, Brisbane, Australia. E-mail: D.Hume{at}cmcb.uq.edu.au

The c-fms gene encodes the receptor for macrophage colony-stimulating factor-1. This gene is expressed selectively in the macrophage cell lineage. Previous studies have implicated sequences in intron 2 that control transcript elongation in tissue-specific and regulated expression of c-fms. Four macrophage-specific deoxyribonuclease I (DNase I)-hypersensitive sites (DHSs) were identified within mouse intron 2. Sequences of these DHSs were found to be highly conserved compared with those in the human gene. A 250-bp region we refer to as the fms intronic regulatory element (FIRE), which is even more highly conserved than the c-fms proximal promoter, contains many consensus binding sites for macrophage-expressed transcription factors including Sp1, PU.1, and C/EBP. FIRE was found to act as a macrophage-specific enhancer and as a promoter with an antisense orientation preference in transient transfections. In stable transfections of the macrophage line RAW264, as well as in clones selected for high- and low-level c-fms mRNA expression, the presence of intron 2 increased the frequency and level of expression of reporter genes compared with those attained using the promoter alone. Removal of FIRE abolished reporter gene expression, revealing a suppressive activity in the remaining intronic sequences. Hence, FIRE is shown to be a key regulatory element in the fms gene.

Key Words: intron • enhancer • transcription • DNase I hypersensitivity




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