The human granulocyte-macrophage colony-stimulating factor gene is autonomously regulated in vivo by an inducible tissue-specific enhancer

Research output: Contribution to journalArticle

Authors

Colleges, School and Institutes

Abstract

The granulocyte-macrophage colony-stimulating factor (GM-CSF) gene is part of a cytokine gene cluster and is directly linked to a conserved upstream inducible enhancer. Here we examined the in vitro and in vivo functions of the human GM-CSF enhancer and found that it was required for the correctly regulated expression of the GM-CSF gene. An inducible DNase I-hypersensitive site appeared within the enhancer in cell types such as T cells, myeloid cells, and endothelial cells that express GM-CSF, but not in nonexpressing cells. In a panel of transfected cells the human GM-CSF enhancer was activated in a tissue-specific manner in parallel with the endogenous gene. The in vivo function of the enhancer was examined in a transgenic mouse model that also addressed the issue of whether the GM-CSF locus was correctly regulated in isolation from other segments of the cytokine gene cluster. After correction for copy number the mean level of human GM-CSF expression in splenocytes from 11 lines of transgenic mice containing a 10.5-kb human GM-CSF transgene was indistinguishable from mouse GM-CSF expression (99% +/- 56% SD). In contrast, a 9.8-kb transgene lacking just the enhancer had a significantly reduced (P = 0.004) and more variable level of activity (29% +/- 89% SD). From these studies we conclude that the GM-CSF enhancer is required for the correct copy number-dependent expression of the human GM-CSF gene and that the GM-CSF gene is regulated independently from DNA elements associated with the closely linked IL-3 gene or other members of the cytokine gene cluster.

Details

Original languageEnglish
Pages (from-to)15097-102
Number of pages6
JournalNational Academy of Sciences. Proceedings
Volume96
Issue number26
Publication statusPublished - 21 Dec 1999

Keywords

  • Animals, Binding Sites, Chromatin, Cytokines, DNA Footprinting, Deoxyribonuclease I, Enhancer Elements, Genetic, Gene Dosage, Gene Expression Regulation, Granulocyte-Macrophage Colony-Stimulating Factor, HeLa Cells, Humans, Interleukin-3, Jurkat Cells, Mice, Mice, Transgenic, Multigene Family, Tissue Distribution, U937 Cells