Stefanovic B, Marzluff W F
Institute of Molecular Biophysics, Tallahassee, FL 32306.
Gene Expr. 1994;4(1-2):1-18.
There are two gene sets for U2 snRNA in the sea urchin L. variegatus, the U2E gene, which is expressed in oogenesis and early embryogenesis and then silenced, and the U2L gene, which is expressed constitutively. There are four major promoter elements found in both U2 snRNA genes; an essential TATA box at -25 to -30, a proximal element (PSE) at -55 required for expression, an element at -100 necessary for maximal expression, and an upstream activating sequence (UAS) necessary for maximal expression. The elements of the U2E and U2L promoters were interchanged and the effect on the temporal pattern of expression determined after microinjection of the genes into sea urchin zygotes. When the U2E PSE element was introduced into the U2L gene, the temporal pattern of expression of the gene was changed to that of the U2E gene. Converting the U2L gene proximal element into the early U2 gene proximal element by altering 14 nucleotides in the promoter also changed the temporal pattern of expression of the U2L gene. Factors that interact with the U2E PSE, detected by a gel mobility shift assay and DNaseI footprinting, were present in blastula but not late gastrula embryos. In contrast, interchanging the -100 element did not greatly affect the temporal pattern of expression, and factors that interact with the U2E gene -100 box were present in both late gastrula and blastula embryos.
在多色紫球海胆中,U2 snRNA有两个基因集,即U2E基因,它在卵子发生和早期胚胎发生过程中表达,然后沉默;还有U2L基因,它持续表达。在这两个U2 snRNA基因中都发现了四个主要的启动子元件;一个位于-25至-30的必需TATA框,一个位于-55的近端元件(PSE),它是表达所必需的,一个位于-100的元件,它是最大表达所必需的,以及一个上游激活序列(UAS),它是最大表达所必需的。将U2E和U2L启动子的元件进行互换,并在将基因显微注射到海胆受精卵后,确定其对表达时间模式的影响。当将U2E PSE元件引入U2L基因时,该基因的表达时间模式改变为U2E基因的模式。通过改变启动子中的14个核苷酸,将U2L基因近端元件转化为早期U2基因近端元件,也改变了U2L基因的表达时间模式。通过凝胶迁移率变动分析和DNaseI足迹法检测到的与U2E PSE相互作用的因子,存在于囊胚期胚胎中,但不存在于原肠胚后期胚胎中。相反,互换-100元件对表达时间模式影响不大,与U2E基因-100框相互作用的因子在原肠胚后期和囊胚期胚胎中均存在。