Lysyl oxidase activates the transcription activity of human collagene III promoter - Possible involvement of Ku antigen
Lysyl oxidase activates the transcription activity of human collagene III promoter - Possible involvement of Ku antigen
复制标题
DOI：
10.1074/jbc.m003362200
复制
发表时间：
2000-11-17
影响因子：
4.8
通讯作者：
Di Donato, A
中科院分区：
文献类型：
--
作者：
Giampuzzi, M;Botti, G;Di Donato, A
赖氨酰氧化酶是一种控制胶原和弹性蛋白成熟的胞外酶，赖氨酰氧化酶和胶原III在纤维化组织中通常显示相似的表达模式。因此，我们研究赖氨酰氧化酶过表达对人COL 3A 1基因启动子活性的影响。我们的结果表明，当COS-7细胞过表达赖氨酰氧化酶的成熟形式时，当通过荧光素酶报告基因检测时，人COL 3A 1启动子的活性平均增加高达12倍。这种效应是特异性的，因为其他启动子不受影响。此外，赖氨酰氧化酶的催化活性可被特异性抑制剂β-氨基丙腈阻断.电泳迁移率变动分析显示，在-101 ~-77区域的结合活性被赖氨酰氧化酶过表达显著增加，该结合被冷探针特异性竞争，并且该区域的突变在转染实验中消除了凝胶阻滞中的结合活性和对COL 3A 1启动子的赖氨酰氧化酶刺激。我们鉴定了其两种组分Ku 80和Ku 70的Ku抗原结合活性。这项研究提出了一种新的协调机制，赖氨酰氧化酶可能控制纤维化的发展。
Lysyl oxidase is an extracellular enzyme that controls the maturation of collagen and elastin, Lysyl oxidase and collagen III often show similar expression patterns in fibrotic tissues. Therefore, we investigated the influence of lysyl oxidase overexpression on the promoter activity of human COL3A1 gene. Our results showed that when COS-7 cells overexpressed the mature form of lysyl oxidase, the activity of the human COL3A1 promoter was increased up to an average of 12 times when tested by luciferase reporter assay. The effect was specific, because other promoters were not affected. Moreover, lysyl oxidase effect was abolished by beta -aminopropionitrile, a specific inhibitor of its catalytic activity. Electrophoretic mobility shift assay showed a binding activity in the region from -101 to -77 that was significantly increased by lysyl oxidase overexpression, The binding was specifically competed by the cold probe, and the mutagenesis of this region abolished both the binding activity in gel retardation and lysyl oxidase stimulation of COL3A1 promoter in transfection experiments. We identified the binding activity as Ku antigen in its two components: Ku80 and Ku70. This study suggests a new coordinated mechanism by which lysyl oxidase might control the development of fibrosis.