Svendsen P, Laursen J, Krogh-Pedersen H, Hjorth J P
Department of Molecular and Structural Biology, Aarhus University, DK-8000 Aarhus C, Denmark.
Nucleic Acids Res. 1998 Jun 1;26(11):2761-70. doi: 10.1093/nar/26.11.2761.
The murine parotid secretory protein (PSP) gene is expressed selectively at high levels in parotid and sublingual salivary glands. Previously, the transcriptional activity of a PSP mini-gene, called Lama, was shown to be dependent on a 1.5 kb region located 3 kb upstream of the transcription start site. Here, functional studies in transgenic mice demonstrate that this proximal regulatory region has properties of a parotid and sublingual gland specific enhancer. Protein-binding experiments identify multiple sequence-specific binding complexes spanning the entire 1.5 kb enhancer region. Several sequence elements bound specifically by parotid and/or sublingual gland nuclear extracts, including consensus binding elements for previously described transcription factors as well as novel binding elements are located in the proximal enhancer region. A deletion analysis of the enhancer region in transgenic mice identified a core sequence of 700 bp. This region contains five elements bound specifically by nuclear proteins isolated from the PSP-expressing parotid and sublingual glands. Two of these elements, denoted parotid gland element I (PGE I) and sublingual gland element I (SLE I), are novel salivary gland specific binding elements, bound uniquely by parotid and sublingual gland nuclear extracts, respectively.
小鼠腮腺分泌蛋白(PSP)基因在腮腺和舌下唾液腺中选择性地高水平表达。此前,一个名为Lama的PSP小基因的转录活性被证明依赖于转录起始位点上游3 kb处的一个1.5 kb区域。在此,对转基因小鼠的功能研究表明,这个近端调控区域具有腮腺和舌下腺特异性增强子的特性。蛋白质结合实验确定了跨越整个1.5 kb增强子区域的多个序列特异性结合复合物。腮腺和/或舌下腺核提取物特异性结合的几个序列元件,包括先前描述的转录因子的共有结合元件以及新的结合元件,位于近端增强子区域。对转基因小鼠中增强子区域的缺失分析确定了一个700 bp的核心序列。该区域包含五个与从表达PSP的腮腺和舌下腺分离的核蛋白特异性结合的元件。其中两个元件,分别称为腮腺元件I(PGE I)和舌下腺元件I(SLE I),是新的唾液腺特异性结合元件,分别被腮腺和舌下腺核提取物独特地结合。