Buono P, de Conciliis L, Olivetta E, Izzo P, Salvatore F
Dipartimento di Biochimica e Biotecnologie Mediche, Facoltà di Medicina e Chirurgia, Università di Napoli Federico II, Italy.
FEBS Lett. 1993 Aug 16;328(3):243-9. doi: 10.1016/0014-5793(93)80936-o.
We investigated the cis-acting sequences involved in the expression of the human aldolase C gene by transient transfections into human neuroblastoma cells (SKNBE). We demonstrate that 420 bp of the 5'-flanking DNA direct at high efficiency the transcription of the CAT reporter gene. A deletion between -420 bp and -164 bp causes a 60% decrease of CAT activity. Gel shift and DNase I footprinting analyses revealed four protected elements: A, B, C and D. Competition analyses indicate that Sp1 or factors sharing a similar sequence specificity bind to elements A and B, but not to elements C and D. Sequence analysis shows a half palindromic ERE motif (GGTCA), in elements B and D. Region D binds a transactivating factor which appears also essential to stabilize the initiation complex.
我们通过瞬时转染人神经母细胞瘤细胞（SKNBE）研究了参与人醛缩酶C基因表达的顺式作用序列。我们证明，5'-侧翼DNA的420 bp高效指导CAT报告基因的转录。-420 bp至-164 bp之间的缺失导致CAT活性降低60%。凝胶迁移和DNase I足迹分析揭示了四个受保护元件：A、B、C和D。竞争分析表明，Sp1或具有相似序列特异性的因子与元件A和B结合，但不与元件C和D结合。序列分析显示，元件B和D中存在一个半回文ERE基序（GGTCA）。区域D结合一种反式激活因子，该因子似乎对稳定起始复合物也至关重要。
