Kuzmin I, Duh F M, Latif F, Geil L, Zbar B, Lerman M I
Laboratory of Immunobiology, NCI-Frederick Cancer Research and Development Center, Maryland 21702-1201, USA.
Oncogene. 1995 Jun 1;10(11):2185-94.
The von Hippel-Lindau (VHL) disease gene is a novel multiple tumor suppressor gene which plays a causal role in the origin of some common cancers including clear cell renal carcinomas and hemangioblastomas of the central nervous system. Here we report the identification of transcription start sites and the promoter of the human VHL gene. The promoter sequence does not contain TATA and CCAAT boxes. Transcription is initiated around a putative SP1 binding site about 60 bp upstream from the first AUG codon in the VHL mRNA. Several putative transcription factor binding sites, notably for nuclear respiratory factor 1 and PAX, were found upstream of the transcription start sites. Promoter-luciferase expression constructs demonstrate, that the promoter is functional when transfected into 293 cells (transformed primary human embryonal kidney cells) and UMRC 6 renal carcinoma cells. Activity is dependent on correct orientation of the promoter. A minimal promoter region of 106 bp was delineated. A set of VHL minigenes, containing the 5' flanking VHL genomic region, was constructed and transfected into UMRC 6 cells. In these cells the level of transcription from the minigenes driven by VHL promoter was comparable with endogenous VHL expression.
冯·希佩尔-林道（VHL）病基因是一种新型的多肿瘤抑制基因，在包括透明细胞肾癌和中枢神经系统血管母细胞瘤在内的一些常见癌症的发生中起因果作用。在此，我们报告了人类VHL基因转录起始位点和启动子的鉴定。启动子序列不含TATA盒和CCAAT盒。转录在VHL mRNA中第一个AUG密码子上游约60 bp处的一个假定的SP1结合位点周围起始。在转录起始位点上游发现了几个假定的转录因子结合位点，特别是核呼吸因子1和PAX的结合位点。启动子-荧光素酶表达构建体表明，该启动子转染到293细胞（转化的原代人胚肾细胞）和UMRC 6肾癌细胞中时具有功能。活性取决于启动子的正确方向。确定了一个106 bp的最小启动子区域。构建了一组包含VHL基因组5'侧翼区域的VHL微型基因，并将其转染到UMRC 6细胞中。在这些细胞中，由VHL启动子驱动的微型基因的转录水平与内源性VHL表达相当。
