Thiel G, Greengard P, Südhof T C
Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, NY 10021.
Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3431-5. doi: 10.1073/pnas.88.8.3431.
Synapsin Ia and synapsin Ib are abundant synaptic vesicle proteins that are derived by differential splicing from a single gene. To identify control elements directing the neuronal expression of synapsins Ia/b, we functionally analyzed the promoter region of the human synapsin I gene. A hybrid gene was constructed containing 2 kilobases of 5' flanking sequence from the synapsin I gene fused to the bacterial gene chloramphenicol acetyltransferase and transfected into 12 different neuronal and nonneuronal cell lines. In general, expression of the chimeric reporter gene showed excellent correlation with endogenous expression of synapsin I in different neuronal cell lines, whereas transcription was low in all nonneuronal cell lines examined. The addition of the simian virus 40 enhancer promoted non-tissue-specific expression. Deletion mutagenesis of the synapsin I promoter revealed the presence of positive and negative sequence elements. A basal (constitutive) promoter that directs reporter gene expression in neuronal and nonneuronal cell lines was mapped to the region -115 to +47. The promoter region from -422 to -22 contains positive elements that upon fusion with the herpes simplex virus thymidine kinase promoter potentiate its transcription in PC12 and neuroblastoma cells but not in Chinese hamster ovary cells.
突触素Ia和突触素Ib是丰富的突触小泡蛋白,它们由单个基因通过差异剪接产生。为了鉴定指导突触素Ia/b神经元表达的调控元件,我们对人突触素I基因的启动子区域进行了功能分析。构建了一个杂合基因,该基因包含来自突触素I基因的2千碱基5'侧翼序列,与细菌氯霉素乙酰转移酶基因融合,并转染到12种不同的神经元和非神经元细胞系中。一般来说,嵌合报告基因的表达与不同神经元细胞系中突触素I的内源性表达具有良好的相关性,而在所检测的所有非神经元细胞系中转录水平较低。添加猿猴病毒40增强子可促进非组织特异性表达。突触素I启动子的缺失诱变揭示了正负序列元件的存在。一个在神经元和非神经元细胞系中指导报告基因表达的基础(组成型)启动子被定位到-115至+47区域。-422至-22的启动子区域包含正向元件,这些元件与单纯疱疹病毒胸苷激酶启动子融合后,可增强其在PC12细胞和神经母细胞瘤细胞中的转录,但在中国仓鼠卵巢细胞中则不然。