Logan J, Falck-Pedersen E, Darnell J E, Shenk T
Agricultural Research Division, American Cyanamid Company, Princeton, NJ 08540.
Proc Natl Acad Sci U S A. 1987 Dec;84(23):8306-10. doi: 10.1073/pnas.84.23.8306.
Sequence elements within the mouse beta maj-globin transcription unit required for efficient termination of transcription by RNA polymerase II have been delineated. To facilitate nascent-chain analysis of termination, the DNA segment in which transcription ceases was introduced into the adenovirus chromosome within its E1A transcription unit. Two beta-globin DNA elements were required to effect efficient termination: an upstream sequence that includes two poly(A) addition signals and a downstream region previously shown to be where RNA synthesis stops. The role of poly(A) addition in termination was established by introduction of several single base pair substitutions into the AATAAA polyadenylylation motifs. These mutations inhibited both polyadenylylation and termination within the beta-globin DNA segment. Therefore, poly(A) addition appears to be a prerequisite for efficient termination.
已经确定了小鼠β-珠蛋白主要转录单位内RNA聚合酶II有效终止转录所需的序列元件。为了便于对终止进行新生链分析,将转录终止的DNA片段引入腺病毒染色体的E1A转录单位内。实现有效终止需要两个β-珠蛋白DNA元件:一个上游序列,其中包括两个聚腺苷酸化信号,以及一个先前显示为RNA合成停止位置的下游区域。通过将几个单碱基对替换引入AATAAA聚腺苷酸化基序,确定了聚腺苷酸化在终止中的作用。这些突变抑制了β-珠蛋白DNA片段内的聚腺苷酸化和终止。因此,聚腺苷酸化似乎是有效终止的先决条件。