Wang W, Klein W H
Department of Biochemistry and Molecular Biology, University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
DNA Cell Biol. 1996 Feb;15(2):133-45. doi: 10.1089/dna.1996.15.133.
The LpS1 beta gene of Lytechinus pictus is activated at the late cleavage stage about 12 hr after fertilization. LpS1 beta transcripts accumulate exclusively in aboral ectoderm lineages. LpS1 beta is thus a classic example of a gene whose expression is tightly controlled both temporally and spatially during early development. Previous studies on the LpS1 beta promoter identified two G-string DNA elements, one proximal and one distal to the LpS1 beta transcriptional start site, which bind to an ectoderm-enriched nuclear factor. In this report, we show that the ectoderm G-string factor binds to a G/C-rich region larger than the G-string itself and that the binding of the G-string factor requires sequences immediately downstream from the G-string. These downstream sequences are essential for full promoter activity. Two regions of 5'-flanking DNA are required for positive control of LpS1 beta, region I from base pairs -762 to -511, and region II, which includes the G/C-rich element, from base pairs -108 to -61. Region I also contains a mesenchyme cell repressor element. The results indicate that LpS1 beta expression is regulated positively in ectoderm cells and negatively in mesenchyme cells. Similar positive and negative control mechanisms regulate the expression of the related Spec genes of Strongylocentrotus purpuratus, but in this gene family the DNA elements are entirely different. We hypothesize that cis-regulatory elements are evolutionarily dynamic and that many different combinations of DNA elements are capable of given rise to aboral ectoderm-specific expression.
刺冠海胆的LpS1β基因在受精后约12小时的卵裂后期被激活。LpS1β转录本仅在反口外胚层谱系中积累。因此,LpS1β是一个典型的例子,说明一个基因的表达在早期发育过程中受到时间和空间上的严格控制。先前对LpS1β启动子的研究确定了两个G串DNA元件,一个位于LpS1β转录起始位点近端,一个位于远端,它们与富含外胚层的核因子结合。在本报告中,我们表明外胚层G串因子与一个比G串本身更大的富含G/C的区域结合,并且G串因子的结合需要G串下游的紧邻序列。这些下游序列对于启动子的完全活性至关重要。LpS1β的正向调控需要5'侧翼DNA的两个区域,区域I从碱基对-762到-511,区域II包括富含G/C的元件,从碱基对-108到-61。区域I还包含一个间充质细胞抑制元件。结果表明,LpS1β在外胚层细胞中受到正向调控,而在间充质细胞中受到负向调控。类似的正向和负向调控机制调节紫海胆相关Spec基因的表达,但在这个基因家族中,DNA元件完全不同。我们推测顺式调控元件在进化上是动态的,并且许多不同的DNA元件组合能够导致反口外胚层特异性表达。