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Identification and analysis of the human hyaluronan synthase 1 gene promoter reveals Smad3- and Sp3-mediated transcriptional induction.
- Source :
-
Matrix biology : journal of the International Society for Matrix Biology [Matrix Biol] 2012 Sep-Oct; Vol. 31 (7-8), pp. 373-9. Date of Electronic Publication: 2012 Nov 01. - Publication Year :
- 2012
-
Abstract
- The ubiquitous mammalian extracellular matrix glycosaminoglycan hyaluronan (HA) plays a pivotal role in the regulation of cell phenotype in fibrosis and scarring. Transforming growth factor-beta 1 (TGF-β1) and interleukin-1 beta (IL-1β) up-regulate hyaluronan synthase (HAS) 1 and HAS2 in dermal fibroblasts and renal proximal tubular epithelial cells, and subsequent HA synthesis regulates cell phenotype. In the present study, we investigated the mechanism of HAS1 transcriptional up-regulation in response to these cytokines. We used 5'-rapid amplification of cDNA ends analysis to identify the 5' end of HAS1 transcripts, resulting in an increase of 26 nucleotides to the HAS1 exon 1 sequence of reference sequence NM&#95;001523. Constitutive luciferase activity of upstream DNA sequences was shown in luciferase reporter assays, but our reporter vector signals were refractory to the addition of TGF-β1 and IL-1β. Using siRNAs to knockdown transcription factor mRNAs, we showed that TGF-β1 up-regulation of HAS1 transcription was mediated via Smad3 but not Smad2, while HAS1 induction by IL-1β was Sp3, not Sp1, dependent.<br /> (Copyright © 2012 Elsevier B.V. All rights reserved.)
- Subjects :
- Base Sequence
DNA Primers genetics
DNA, Complementary genetics
Electrophoresis, Agar Gel
Gene Expression Regulation physiology
Humans
Hyaluronan Synthases
Interleukin-1beta metabolism
Luciferases
Molecular Sequence Data
Nucleic Acid Amplification Techniques
RNA Interference
RNA, Small Interfering genetics
Reverse Transcriptase Polymerase Chain Reaction
Sequence Analysis, DNA
Extracellular Matrix Proteins genetics
Gene Expression Regulation genetics
Glucuronosyltransferase genetics
Promoter Regions, Genetic genetics
Smad3 Protein metabolism
Sp3 Transcription Factor metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1569-1802
- Volume :
- 31
- Issue :
- 7-8
- Database :
- MEDLINE
- Journal :
- Matrix biology : journal of the International Society for Matrix Biology
- Publication Type :
- Academic Journal
- Accession number :
- 23123404
- Full Text :
- https://doi.org/10.1016/j.matbio.2012.10.002