Dataset Information

Cloning and characterization of the human integrin ?6 gene promoter.

ABSTRACT: The integrin ?6 (ITGB6) gene, which encodes the limiting subunit of the integrin ?v?6 heterodimer, plays an important role in wound healing and carcinogenesis. The mechanism underlying ITGB6 regulation, including the identification of DNA elements and cognate transcription factors responsible for basic transcription of human ITGB6 gene, remains unknown. This report describes the cloning and characterization of the human ITGB6 promoter. Using 5'-RACE (rapid amplification of cDNA ends) analysis, the transcriptional initiation site was identified. Promoter deletion analysis identified and functionally validated a TATA box located in the region -24 to -18 base pairs upstream of the ITGB6 promoter. The regulatory elements for transcription of the ITGB6 gene were predominantly located -289 to -150 from the ITGB6 promoter and contained putative binding sites for transcription factors such as STAT3 and C/EBP?. Using chromatin immunoprecipitation assays, this study has demonstrated, for the first time, that transcription factors STAT3 and C/EBP? are involved in the positive regulation of ITGB6 transcription in oral squamous cell carcinoma cells. These findings have important implications for unraveling the mechanism of abnormal ITGB6 activation in tissue remodeling and tumorigenesis.

SUBMITTER: Xu M

PROVIDER: S-EPMC4376883 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Cloning and characterization of the human integrin β6 gene promoter.

Xu Mingyan M Chen Xihe X Yin Hao H Yin Liqin L Liu Fan F Fu Yucai Y Yao Jiangwu J Deng Xiaoling X

PloS one 20150327 3

The integrin β6 (ITGB6) gene, which encodes the limiting subunit of the integrin αvβ6 heterodimer, plays an important role in wound healing and carcinogenesis. The mechanism underlying ITGB6 regulation, including the identification of DNA elements and cognate transcription factors responsible for basic transcription of human ITGB6 gene, remains unknown. This report describes the cloning and characterization of the human ITGB6 promoter. Using 5'-RACE (rapid amplification of cDNA ends) analysis, t ...[more]

PMID: 25816241

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Project description:Î±9b1 is the most recent addition to the integrin family of membrane-bound receptors and consequently remains the one which is the least characterized. To better understand how transcription of the human gene encoding the Î±9 subunit is regulated at the molecular level, we cloned the Î±9 promoter and characterized the regulatory elements that are required to ensure its transcription. Transfection of Î±9 promoter/CAT recombinant plasmids in primary cultured cells and uveal melanoma cell lines demonstrated the presence of both negative and positive regulatory elements along the Î±9 promoter and positioned the basal Î±9 promoter to within 118 bp from the Î±9 mRNA start site. In vitro DNaseI footprinting and in vivo ChIP analyses demonstrated the binding of the transcription factors Sp1/Sp3, c-Myb and NFI to the most upstream Î±9 negative regulatory element. The transcription factors Sp1/Sp3 and NFI were found to bind the basal Î±9 promoter individually but Sp1 binding clearly predominates when both transcription factors are present in the same extract. Most of all, addition of tenascin-C (TNC), the ligand of Î±9Î²1, to the tissue culture plates prior to cell seeding increased Î±9 transcription whereas it simultaneously decreased expression of the Î±5 integrin subunit gene. This dual regulatory action of TNC on the transcription of the Î±9 and Î±5 genes suggests that both these integrins must work together to appropriately regulate cell adhesion, migration and differentiation that are hallmarks of tissue wound healing. Primary cultures of human corneal epithelial cells (HCECs; number of replicates: 8), human skin epithelial cells (HSEC; number of replicates: 4), human corneal fibroblast cells (HCFCs; number of replicates: 3), human skin fibroblast cells (HSFCs; number of replicates: 3), human uveal melanocytes (UVM; number of replicates: 3), and various humal uveal melanoma cell lines (T115, T142 and T143; number of replicates: 2-3) were analyzed by gene profiling on microarrays at low culture passages (between passage 2 to 7). HCECs were also cultured on either BSA (number of replicates: 2) or on tenascin-C (number of replicates: 2) and their transcriptome analyzed by gene profiling.

Dataset Information

Cloning and characterization of the human integrin ?6 gene promoter.

Publications

Cloning and characterization of the human integrin β6 gene promoter.

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