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Isolation and characterization of the platelet‐derived growth factor beta receptor promoter
Author(s) -
Shinbrot Eve,
Liao Xiang,
Williams Lewis T.
Publication year - 1997
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/(sici)1097-0177(199702)208:2<211::aid-aja8>3.0.co;2-l
Subject(s) - biology , reporter gene , microbiology and biotechnology , chloramphenicol acetyltransferase , gene expression , promoter , gene , regulation of gene expression , transcription factor , endogeny , regulatory sequence , genetics , endocrinology
The PDGFβr gene has been implicated in many physiological processes including development and wound healing. Aberrant expression of the receptor is seen in many pathological conditions such as atherosclerosis and inflammatory diseases. To study the mechanisms of PDGFβr regulation, we identified the regulatory regions of the gene. We have cloned and characterized the promoter region of the platelet‐derived growth factor beta receptor (PDGFβr). We isolated a 4.5 Kb genomic fragment which confers PDGFβr tissue‐specific promoter activity. This fragment can direct transcription of a luciferase reporter gene in a cell‐specific manner which correlates well with the known pattern of expression of the PDGFβr. The specificity of this clone was demonstrated by its high activity in NIH 3T3 fibroblasts and lack of activity in N‐MUNG epithelial cells, a pattern that parallels the expression of the endogenous PDGFβr. We have defined a 614 bp region encompassing the 5′ untranslated region of the gene which includes the basal promoter region. We generated transgenic mice that carry the chloramphenicol acetyltransferase (CAT) reporter gene under the control of the 4.5 Kb promoter. The expression pattern of the reporter gene was compared to that of the endogenous PDGFβr gene. The promoter was able to direct reporter gene expression with the same temporal and spatial pattern as the endogenous PDGFβr. The most prominent expression was in condensing mesenchyme of developing blood vessels, bone and tissues adjacent to epithelium. We conclude that this clone contains the regulatory regions sufficient to direct expression of the PDGFβr. The further analysis of this promoter will help elucidate the transcriptional regulation of expression of the PDGFβr, and provide a useful tool for directing expression of heterologous genes. Dev. Dyn. 208:211–219, 1997. © 1997 Wiley‐Liss, Inc.