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Identification and analysis of the promoter region of the human hyaluronan synthase 2 gene

Monslow, Jamie, Williams, John David, Guy, Carol, Price, Iain Kelsey, Craig, Kathrine Jane, Williams, Hywel John ORCID:, Williams, Nigel Melville ORCID:, Martin, John, Coleman, Sharon Louise, Topley, Nicholas, Spicer, Andrew P., Buckland, Paul Robert, Davies, Malcolm and Bowen, Timothy ORCID: 2004. Identification and analysis of the promoter region of the human hyaluronan synthase 2 gene. The Journal of Biological Chemistry 279 (20) , pp. 20576-20581. 10.1074/jbc.M312666200

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Hyaluronan (HA) is a linear glycosaminoglycan of the vertebrate extracellular matrix that is synthesized at the plasma membrane by the HA synthase (HAS) enzymes HAS1, -2 and -3. The regulation of HA synthesis has been implicated in a variety of extracellular matrix-mediated and pathological processes, including renal fibrosis. We have recently described the genomic structures of each of the human HAS genes. In the present study, we analyzed the HAS2 promoter region. In 5′-rapid amplification of cDNA ends analysis of purified mRNA from human renal epithelial proximal tubular cells, we detected an extended sequence for HAS2 exon 1, relocating the transcription initiation site 130 nucleotides upstream of the reference HAS2 mRNA sequence, GenBank™ accession number NM_005328. A luciferase reporter gene assay of nested fragments spanning the 5′ terminus of NM_005328 demonstrated the constitutive promoter activity of sequences directly upstream of the repositioned transcription initiation site but not of the newly designated exonic nucleotides. Using reverse transcription-PCR, expression of this extended HAS2 mRNA was demonstrated in a variety of human cell types, and orthologous sequences were detected in mouse and rat kidney. Alignment of human, murine, and equine genomic DNA sequences upstream of the repositioned HAS2 exon 1 provided evidence for the evolutionary conservation of specific transcription factor binding sites. The location of the HAS2 promoter will facilitate analysis of the transcriptional regulation of this gene in a variety of pathological contexts as well as in developmental models in which HAS2 null animals have an embryonic lethal phenotype.

Item Type: Article
Date Type: Publication
Status: Published
Schools: MRC Centre for Neuropsychiatric Genetics and Genomics (CNGG)
Systems Immunity Research Institute (SIURI)
Neuroscience and Mental Health Research Institute (NMHRI)
Subjects: R Medicine > R Medicine (General)
Publisher: American Society for Biochemistry and Molecular Biology
ISSN: 1083-351X
Last Modified: 01 Dec 2022 09:42

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