Identification and Analysis of the Promoter Region of the Human Hyaluronan Synthase 2 Gene
| Autor: | Andrew P. Spicer, Jamie Monslow, Malcolm Davies, Timothy Bowen, Nigel Williams, Iain Kelsey Price, Hywel Williams, John D. Williams, Sharon Louise Coleman, John Martin, Paul Robert Buckland, Kathrine Jane Craig, Carol Guy, Nicholas Topley |
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| Rok vydání: | 2004 |
| Předmět: |
endocrine system
Molecular Sequence Data Biology Kidney Biochemistry Cell Line Conserved sequence Mice Exon Sequence Homology Nucleic Acid Complementary DNA Tumor Cells Cultured Transcriptional regulation Animals Humans Glucuronosyltransferase Promoter Regions Genetic Molecular Biology Gene DNA Primers HAS1 Expressed Sequence Tags Base Sequence Reverse Transcriptase Polymerase Chain Reaction Promoter Cell Biology Molecular biology Rats DNA binding site Hyaluronan Synthases Sequence Alignment |
| Zdroj: | Journal of Biological Chemistry. 279:20576-20581 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m312666200 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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