Localization of silencer and enhancer elements in the human type X collagen gene

Beier, Frank, Vornehm, Silvia, Pöschl, Ernst, von der Mark, Klaus and Lammi, Mikko J. (1997) Localization of silencer and enhancer elements in the human type X collagen gene. Journal of Cellular Biochemistry, 66 (2). pp. 210-8. ISSN 0730-2312

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Abstract

Collagen type X is a short, network-forming collagen expressed temporally and spatially tightly controlled in hypertrophic chondrocytes during endochondral ossification. Studies on chicken chondrocytes indicate that the regulation of type X collagen gene expression is regulated at the transcriptional level. In this study, we have analyzed the regulatory elements of the human type X collagen (Col10a1) by reporter gene constructs and transient transfections in chondrogenic and nonchondrogenic cells. Four different promoter fragments covering up to 2,864 bp of 5'-flanking sequences, either including or lacking the first intron, were linked to luciferase reporter gene and transfected into 3T3 fibroblasts, HT1080 fibrosarcoma cells, prehypertrophic chondrocytes from the resting zone, hypertrophic chondrocytes, and chondrogenic cell lines. The results indicated the presence of three regulatory elements in the human Col10a1 gene besides the proximal promoter. First, a negative regulatory element located between 2.4 and 2.8 kb upstream of the transcription initiation site was active in all nonchondrogenic cells and in prehypertrophic chondrocytes. Second, a positive, but also non-tissue-specific positive regulatory element was present in the first intron. Third, a cell-type-specific enhancer element active only in hypertrophic chondrocytes was located between -2.4 and -0.9 kb confirming a previous report by Thomas et al. [(1995): Gene 160:291-296]. The enhancing effect, however, was observed only when calcium phosphate was either used for transfection or included in the culture medium after lipofection. These findings demonstrate that the rigid control of human Col10a1 gene expression is achieved by both positive and negative regulatory elements in the gene and provide the basis for the identification of factors binding to those elements.

Item Type:	Article
Uncontrolled Keywords:	animals,calcium phosphates,cartilage, articular,cattle,cell line,collagen,enhancer elements, genetic,gene expression regulation,hypertrophy,introns,mice,promoter regions, genetic,transfection
Faculty \ School:	Faculty of Science > School of Biological Sciences
UEA Research Groups:	Faculty of Science > Research Groups > Cells and Tissues
Depositing User:	Pure Connector
Date Deposited:	13 Jan 2016 12:01
Last Modified:	19 Apr 2023 23:45
URI:	https://ueaeprints.uea.ac.uk/id/eprint/56287
DOI:	10.1002/(SICI)1097-4644(19970801)66:2<210::AID-JCB8>3.0.CO;2-T

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