Romanelli Maria Grazia, Lorenzi Pamela, Morandi Carlo
Department of Mother and Child, Biology and Genetics, University of Verona, Strada le Grazie 8, 37134 Verona, Italy.
Gene. 2005 Aug 15;356:11-8. doi: 10.1016/j.gene.2005.04.031.
Neural polypyrimidine tract-binding protein nPTB, originally identified as the neuronal counterpart of the hnRNPI/PTB protein, is an RNA binding protein involved into tissue-specific alternative splicing regulation. Here we describe the functional characterization of the promoter sequence of nPTB in HeLa and neuroblastoma cells. By means of genomic sequence analysis we have isolated and cloned a 1587-base pair region upstream the human nPTB coding region. The nPTB proximal promoter, although rich in G+C content and presenting putative binding sites for the transcription factors Sp1, NF-1, NF-kB and Oct-1, lacks a typical TATA box. Luciferase transient expression assays using deletion mutants have identified the proximal promoter region at -125 relative to the transcription start site. Alignment of human, murine and chimpanzee genomic sequences upstream the nPTB exon 1 has provided evidences for the evolutionary conservation of specific transcription factor binding sites.
神经多嘧啶序列结合蛋白nPTB最初被鉴定为hnRNPI/PTB蛋白的神经元对应物,是一种参与组织特异性可变剪接调控的RNA结合蛋白。在此,我们描述了nPTB启动子序列在HeLa细胞和神经母细胞瘤细胞中的功能特性。通过基因组序列分析,我们分离并克隆了人类nPTB编码区上游1587个碱基对的区域。nPTB近端启动子虽然富含G+C含量,并呈现出转录因子Sp1、NF-1、NF-kB和Oct-1的假定结合位点,但缺乏典型的TATA框。使用缺失突变体的荧光素酶瞬时表达分析确定了相对于转录起始位点-125处的近端启动子区域。nPTB外显子1上游人类、小鼠和黑猩猩基因组序列的比对为特定转录因子结合位点的进化保守性提供了证据。