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Functional Characterization of the Human Biglycan 5′‐Flanking DNA and Binding of the Transcription Factor c‐Krox

Functional Characterization of the Human Biglycan 5′‐Flanking DNA and Binding of the... The transcriptional regulation of human biglycan expression under normal and pathological conditions was studied. The 5′‐flanking regions of the human and mouse genes were isolated and analyzed; the two promoter regions share 81% identity. Both promoters are without a TATA and CAT box and contain multiple Sp1 sites. Human dermal fibroblasts were transiently transfected with progressive deletional human biglycan 5′‐flanking DNA‐CAT constructs, and a significant variation in activity among the individual constructs was found. A small deletion in several cases caused a more than 2‐fold increase or decrease in promoter activity, thereby mapping the target sites for repressors or activators. Human biglycan expression is reduced in females with Ullrich‐Turner syndrome (45,X) and increased in individuals with supernumerary sex chromosomes, and it has been speculated that biglycan plays a role in the short stature phenotype of Turner syndrome. Analysis of the transcriptional regulation of biglycan in individuals with sex chromosome anomalies showed that a −262 to −218 region of the biglycan promoter was differentially regulated. This region was extensively analyzed by DNAse footprinting and electrophoretic mobility shift assays, and a putative binding site for the transcription factor c‐Krox was discovered. The binding of c‐Krox to a site located at approximately −248 to −230 in the human biglycan promoter was confirmed by using extracts from COS cells expressing recombinant human c‐Krox. The expression of c‐Krox in bone was then examined by reverse‐transcribed polymerase chain reaction and Northern blotting analysis; an ∼3.4 kb transcript was detected in primary osteoblastic cells, in MG–63 cells, and in human bone marrow stromal cells. This is the first detection of c‐Krox in bone cells, and it suggests that c‐Krox, like another member of the Krox family, Krox–20, might play a regulatory role in bone. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Bone and Mineral Research Oxford University Press

Functional Characterization of the Human Biglycan 5′‐Flanking DNA and Binding of the Transcription Factor c‐Krox

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References (34)

Publisher
Oxford University Press
Copyright
Copyright © 1997 ASBMR
ISSN
0884-0431
eISSN
1523-4681
DOI
10.1359/jbmr.1997.12.12.2050
pmid
9421237
Publisher site
See Article on Publisher Site

Abstract

The transcriptional regulation of human biglycan expression under normal and pathological conditions was studied. The 5′‐flanking regions of the human and mouse genes were isolated and analyzed; the two promoter regions share 81% identity. Both promoters are without a TATA and CAT box and contain multiple Sp1 sites. Human dermal fibroblasts were transiently transfected with progressive deletional human biglycan 5′‐flanking DNA‐CAT constructs, and a significant variation in activity among the individual constructs was found. A small deletion in several cases caused a more than 2‐fold increase or decrease in promoter activity, thereby mapping the target sites for repressors or activators. Human biglycan expression is reduced in females with Ullrich‐Turner syndrome (45,X) and increased in individuals with supernumerary sex chromosomes, and it has been speculated that biglycan plays a role in the short stature phenotype of Turner syndrome. Analysis of the transcriptional regulation of biglycan in individuals with sex chromosome anomalies showed that a −262 to −218 region of the biglycan promoter was differentially regulated. This region was extensively analyzed by DNAse footprinting and electrophoretic mobility shift assays, and a putative binding site for the transcription factor c‐Krox was discovered. The binding of c‐Krox to a site located at approximately −248 to −230 in the human biglycan promoter was confirmed by using extracts from COS cells expressing recombinant human c‐Krox. The expression of c‐Krox in bone was then examined by reverse‐transcribed polymerase chain reaction and Northern blotting analysis; an ∼3.4 kb transcript was detected in primary osteoblastic cells, in MG–63 cells, and in human bone marrow stromal cells. This is the first detection of c‐Krox in bone cells, and it suggests that c‐Krox, like another member of the Krox family, Krox–20, might play a regulatory role in bone.

Journal

Journal of Bone and Mineral Research – Oxford University Press

Published: Dec 4, 2009

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