CYCLING OF RIBONUCLEIC-ACID POLYMERASE TO PRODUCE OLIGONUCLEOTIDES DURING INITIATION INVITRO AT THE LAC UV5 PROMOTER
CYCLING OF RIBONUCLEIC-ACID POLYMERASE TO PRODUCE OLIGONUCLEOTIDES DURING INITIATION INVITRO AT THE LAC UV5 PROMOTER
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DOI：
10.1021/bi00555a023
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发表时间：
1980-01-01
期刊：
影响因子：
2.9
通讯作者：
GRALLA, JD
中科院分区：
文献类型：
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作者：
CARPOUSIS, AJ;GRALLA, JD
AGamemnon J.Carpousis和Jay D.Gralla摘要：高分辨率凝胶电泳法已被用来检测和定量乳糖操纵子(Lac)UV5启动子在体外转录时产生的启动子特异性寡核苷酸。溶解产物为不同长度的RNA物种，对应于起始的lac mRNA序列。定量表明，每个预引发复合体可以形成许多寡核苷酸，包括长达六核苷酸的物种。在肝素(一种选择性的游离RNA聚合酶抑制剂)存在的情况下，合成过程不会使酶解离，这一点从合成水平上得到了证明。因此，RNA聚合酶在体外循环这个启动子，反复产生寡核苷酸。一般来说，当所有四种底物的总浓度增加时，或当添加缺失的核苷三磷酸底物时，寡核苷酸的产率降低。然而，寡核苷酸合成在所有测试条件下都是有效的。
Agamemnon J. Carpousis* and Jay D. Gralla abstract: High-resolution gel electrophoresis has been used to detect and quantitate promoter-specific oligonucleotides produced duringinitiation of transcriptionin vitro atthe lactose operon (lac) UV5 promoter. Theresolved products are RNA species of various lengths which correspond to the initial lac mRNA sequence. Quantitation shows that many oligo-nucleotides can be formed per preinitiation complex, including species as long as hexanucleotide. Synthesis occurs without dissociation of the enzyme, as evidenced by levels of synthesis in the presence of heparin, a selective inhibitor of free RNA polymerase. Thus, RNA polymerase cycles at this promoter in vitro producing oligonucleotides reiteratively. In general, the yield of oligonucleotides decreases when the total con-centration of all four substrates is increased or when a missing nucleoside triphosphate substrate is added. Nevertheless, oligonucleotide synthesispersists under all conditions tested.