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The microsatellite, macrophage migration inhibitory factor -794, may influence gene expression in human mononuclear
S E L Temple1, K Y Cheong, P Price
1School of Medicine and Pharmacology (RPH Unit), University of Western Australia, Crawley, Australia. stemple@meddent.uwa.edu.au
Abstract:
Polymorphisms within the gene encoding macrophage migration inhibitory factor (MIF) have been associated with susceptibility to inflammatory diseases such as rheumatoid arthritis and increased risk of developing sepsis. We investigated the effects of the MIF-173G>C polymorphism and the MIF-794 CATT microsatellite on MIF expression. These are in moderate linkage disequilibrium. Mononuclear cells from healthy donors were stimulated with bacterial pathogens associated with sepsis (Streptococcus pneumoniae or Escherichia coli ). MIF mRNA and protein levels were measured by real-time polymerase chain reaction and ELISA, respectively. Carriage of the C allele of MIF-173G>C or the 7 CATT repeat of the MIF-794 microsatellite correlated with lower basal and stimulated MIF mRNA levels. However, levels of intracellular and extracellular MIF protein were similar. This discordance between effects on MIF mRNA and protein was not explained by differential effects of genotype on stability of MIF mRNA (detected by actinomycin D mRNA chase). Gel shift assays revealed no differences in the profile of nuclear proteins from mononuclear cells bound by the G and C alleles of MIF-173G>C, but alleles at the microsatellite marker showed differential binding. Our data suggest that the MIF-794 CATT microsatellite influences transcription by differential binding of nuclear transcription factors. This may impact on inflammatory processes.
Insights
Genetic variations in the macrophage migration inhibitory factor (MIF) gene, specifically the MIF-173G>C polymorphism and MIF-794 CATT microsatellite, influence MIF mRNA levels but not protein. The microsatellite appears to affect transcription.
Area of Science:
- Immunogenetics
- Molecular biology
- Inflammation research
Background:
- Polymorphisms in the macrophage migration inhibitory factor (MIF) gene are linked to inflammatory diseases like rheumatoid arthritis and sepsis susceptibility.
- Investigating genetic variants' impact on MIF expression is crucial for understanding disease pathogenesis.
Purpose of the Study:
- To examine the influence of the MIF-173G>C polymorphism and the MIF-794 CATT microsatellite on macrophage migration inhibitory factor (MIF) expression.
- To elucidate the mechanisms underlying the relationship between these genetic markers and MIF levels.
Main Methods:
- Mononuclear cells from healthy donors were stimulated with Streptococcus pneumoniae or Escherichia coli.
- MIF mRNA and protein levels were quantified using real-time polymerase chain reaction and ELISA.
- mRNA stability was assessed via actinomycin D mRNA chase, and nuclear protein binding was analyzed using gel shift assays.
Main Results:
- Carriage of the MIF-173G>C 'C' allele or the MIF-794 CATT 7-repeat microsatellite correlated with reduced basal and stimulated MIF mRNA levels.
- Despite effects on mRNA, intracellular and extracellular MIF protein levels remained similar across genotypes.
- Gel shift assays indicated differential nuclear transcription factor binding at the MIF-794 microsatellite, but not the MIF-173G>C polymorphism.
Conclusions:
- The MIF-794 CATT microsatellite likely influences MIF gene transcription through differential binding of nuclear factors.
- The observed discordance between MIF mRNA and protein levels suggests post-transcriptional regulation or other complex mechanisms.
- These findings may have implications for understanding the role of MIF genetic variations in inflammatory processes and sepsis.
