Functional polymorphism and differential regulation of CYSLTR1 transcription in human airway smooth muscle and monocytes.
Cysteinyl leukotrienes play an important role in the pathophysiology of many inflammatory disorders, including asthma. The aim of this study was to characterize the mechanisms underlying transcriptional regulation of the human cysteinyl leukotriene receptor 1 (hCYSLTR1) gene. 5'RACE was performed on human airway smooth muscle (HASM) and peripheral blood mononuclear cells. A1128-bp region of the hCYSLTR1 main putative promoter was screened for polymorphisms by sequencing of 48 individuals. Luciferase reporter gene assays were performed using fragments of the core promoter (232 bp to 1128 bp) in HASM and THP1 cells. Three hCYSLTR1 transcripts were found, one representing 90% of all messenger RNA identified. The genomic location of the transcription start sites suggested there are two putative hCYSLTR1 promoters. The majority of the transcriptional activity of the main putative promoter was detected between -232 and -679 bp. Four singlenucleotide polymorphisms in strong linkage disequilibrium were found in the region studied: -561 (rs7066737), -642 (rs2806489), -781 (rs2637204), and -940 (rs321029), with three haplotypes observed. In THP1 cells, the G allele (-642) caused a twofold decrease in luciferase expression compared to the Aallele. These data suggest that the majority of hCYSLTR1 transcripts in HASM and monocytes arise from a single promoter located immediately upstream of the 5\' untranslated region, although rarer transcripts can also occur. This study also raises the possibility that cell-type-dependent differences in transcriptional activity caused by the presence of specific haplotypes within the main CYSLTR1 promoter may be a predictor of disease risk or treatment response.
Duroudier
Nathalie P
NP
Division of Therapeutics and Molecular Medicine, Queen's Medical Centre, Nottingham, NG7 2UH, UK. msxnd1@nottingham.ac.uk
Sayers
Ian
I
Castagna
Charlotte Carabott
CC
Fenech
Anthony G
AG
Halapi
Eva
E
Swan
Caroline
C
Hall
Ian P
IP
D000483:Alleles; D017398:Alternative Splicing; D005786:Gene Expression Regulation; D006239:Haplotypes; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D015289:Leukotrienes / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D008957:Models, Genetic; D009000:Monocytes / Q000378:metabolism*; D009130:Muscle, Smooth / Q000378:metabolism*; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic; D018077:Receptors, Leukotriene / Q000378:metabolism*; D012306:Risk; D014132:Trachea / Q000378:metabolism*; D014158:Transcription, Genetic
D015289:Leukotrienes; D008565:Membrane Proteins; D018077:Receptors, Leukotriene; C083169:leukotriene D4 receptor