SIMIAN VIRUS-40 MAJOR LATE PROMOTER - AN UPSTREAM DNA-SEQUENCE REQUIRED FOR EFFICIENT INVITRO TRANSCRIPTION
SIMIAN VIRUS-40 MAJOR LATE PROMOTER - AN UPSTREAM DNA-SEQUENCE REQUIRED FOR EFFICIENT INVITRO TRANSCRIPTION
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DOI：
10.1128/mcb.4.1.133
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发表时间：
1984-01-01
影响因子：
5.3
通讯作者：
SALZMAN, NP
中科院分区：
文献类型：
--
作者：
BRADY, J;RADONOVICH, M;SALZMAN, NP
报道了一个11个碱基的DNA序列，5“-G-G-T-A-C-C-T-A-A-C-C-3”（SV 40图谱位置294-304），它在体外和体内控制SV 40晚期RNA表达中是重要的。鉴定了SV 40晚期启动子的另一个结构域。通过核酸酶BAL 31处理制备一系列具有从SV 40图谱位置0-300延伸的缺失的突变体。然后分析克隆的模板在Manley体外转录系统中晚期SV 40 RNA表达的效率和准确性。除了图谱位置300附近的启动子结构域之外，在核苷酸位置74和95之间存在晚期SV 40 RNA有效表达所需的必需DNA序列。该SV 40 DNA序列中包括6个GGGCGG SV 40重复序列中的2个和一个11个核苷酸的片段，该片段与组蛋白H2 A基因在体外和体内有效表达所需的上游序列具有很强的同源性。该上游启动子序列支持转录具有相同的效率，即使当它被移动72个核苷酸更接近主要的晚期帽位点。体外启动子竞争分析表明，上游启动子序列，独立于294-304启动子元件，能够结合SV 40晚期基因转录所需的聚合酶-转录因子。在核苷酸325处控制RNA起始特异性的DNA序列位于图谱位置294的下游。
An 11-base DNA sequence, 5''-G-G-T-A-C-C-T-A-A-C-C-3'' (SV40 map position 294-304), which is important in the control of SV40 late RNA expression in vitro and in vivo is reported. The identification of another domain of the SV40 late promoter, was identified. A series of mutants with deletions extending from SV40 map position 0-300 was prepared by nuclease BAL 31 treatment. The cloned templates were then analyzed for efficiency and accuracy of late SV40 RNA expression in the Manley in vitro transcription system. In addition to the promoter domain near map position 300, there are essential DNA sequences between nucleotide positions 74 and 95 that are required for efficient expression of late SV40 RNA. Included in this SV40 DNA sequence were 2 of the 6 GGGCGG SV40 repeat sequences and an 11-nucleotide segment which showed strong homology with the upstream sequences required for the efficient in vitro and in vivo expression of the histone H2A gene. This upstream promoter sequence supported transcription with the same efficiency even when it was moved 72 nucleotides closer to the major late cap site. In vitro promoter competition analysis demonstrated that the upstream promoter sequence, independent of the 294-304 promoter element, is capable of binding polymerase-transcription factors required for SV40 late gene transcription. DNA sequences which control the specificity of RNA initiation at nucleotide 325 lie downstream of map position 294.