Grover J, Roughley P J
Genetics Unit, Shriners Hospital for Children, 1529 Cedar Avenue, Montreal, Quebec, Canada H3G 1A6.
Biochem J. 1998 Nov 15;336 ( Pt 1)(Pt 1):77-82. doi: 10.1042/bj3360077.
The 5'-flanking region of the human proline/arginine-rich end leucine-rich repeat protein (PRELP) gene has been characterized for both promoter and repressor activity by using a variety of reporter gene constructs and transient transfection into chondrocytes or fibroblasts. The human PRELP gene lacks a TATA box, and in its absence a Sp1-binding site residing 29 bp upstream of the transcription start site is essential for initiating gene expression. In contrast, an Ets-binding site residing 497 bp upstream of the transcription start site can lead to the repression of gene expression. The analysis of nuclear proteins by gel retardation studies with the repressor element identified a common protein, presumably an Ets family member, present in neonatal chondrocytes and skin fibroblasts that do not express the PRELP gene. The factor was not detected in nuclear protein preparations from adult chondrocytes in which the PRELP gene is expressed.
通过使用多种报告基因构建体并将其瞬时转染到软骨细胞或成纤维细胞中,对人富含脯氨酸/精氨酸的富含亮氨酸重复蛋白(PRELP)基因的5'侧翼区域进行了启动子和阻遏物活性的表征。人PRELP基因缺乏TATA盒,在其缺失的情况下,位于转录起始位点上游29 bp处的Sp1结合位点对于启动基因表达至关重要。相反,位于转录起始位点上游497 bp处的Ets结合位点可导致基因表达的抑制。通过对阻遏元件进行凝胶阻滞研究来分析核蛋白,结果发现一种常见蛋白,可能是Ets家族成员,存在于不表达PRELP基因的新生软骨细胞和皮肤成纤维细胞中。在表达PRELP基因的成年软骨细胞核蛋白制剂中未检测到该因子。