Abstract
Background
Cysteinyl leukotrienes (LT) are mediators involved in inflammatory and allergic disorders. LTC4 synthase catalyzes the first committed step in the synthesis of these inflammatory mediators, and its cellular distribution appears to be unique.
Materials and Methods
A human genomic library was screened by polymerase chain reaction (PCR) with primers that were designed based on the reported cDNA sequence for the LTC4 synthase gene. The gene was identified in one clone by Southern blotting of restriction enzyme digests, subcloning of fragments containing regions of interest, and DNA sequencing of these sub-clones. The transcription initiation site was detennined by primer extension analysis. Chromosome location was determined by fluorescent in situ hybridization and screening of somatic cell hybrids by PCR.
Results
The LTC4 synthase gene is ∼2.5 kb in length, consisting of five exons (136, 100, 71, 82, and 257 bp, respectively) and four introns (1,447, 102, 84, and 230 bp, respectively). Transcription initiation occurs at a single site 78 bp upstream of the coding region. The 5′-flanking region contains neither a TATA nor a CAAT box. The first 1 kb of the 5′-flanking region, however, contains putative DNA binding motifs for SP-1, AP-1, AP-2, ets factors, and CREB/ATF. A STAT binding motif is present in the first intron. The LTC4 synthase gene is located in the distal region of the long arm of chromosome 5 in 5q35.
Conclusions
The LTC4 synthase gene does not contain elements of a typical regulated gene and may therefore contain novel regulatory elements. This gene is also located in a region on chromosome 5 that appears to play a role in allergic and inflammatory disorders, such as asthma.
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Acknowledgments
The authors would like to acknowledge helpful discussions with Dr. Christopher K. Glass. This work was supported in part by a grant-in-aid from the American Heart Association of California, a grant from the Merit Review Board of the Department of Veterans Affairs, and a Career Investigator Award from the American Lung Association (TDB).
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The nucleotide sequence reported in this paper has been deposited in the GenBank database (accession no. U62025).
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Bigby, T.D., Hodulik, C.R., Arden, K.C. et al. Molecular Cloning of the Human Leukotriene C4 Synthase Gene and Assignment to Chromosome 5q35. Mol Med 2, 637–646 (1996). https://doi.org/10.1007/BF03401647
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DOI: https://doi.org/10.1007/BF03401647