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在不含TATA框的腺病毒2型IVa2基因上进行体外转录的精确起始。

In vitro accurate initiation of transcription on the adenovirus type 2 IVa2 gene which does not contain a TATA box.

作者信息

Matsui T

出版信息

Nucleic Acids Res. 1982 Nov 25;10(22):7089-101. doi: 10.1093/nar/10.22.7089.

DOI:10.1093/nar/10.22.7089
PMID:6296765
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC326990/
Abstract

In vitro initiation of transcription has been studied on the IVa2 gene of the adenovirus type 2 ( Ad2 ) which is expressed at intermediate time after infection. This gene does not contain a TATA box around 30 bp upstream from the cap site. By analyses of the in vitro runoff products by size and by single strand nuclease protection mapping, it was concluded that the in vitro initiation occurred accurately at two in vivo cap sites with low but detectable efficiency. By using cloned IVa2 DNA, which is free from the major late promoter, it was suggested that the low efficiency of the in vitro initiation on the IVa2 gene may not be due to close location of the two genes but may rather reflect the efficiency of the promoter itself. The cell-free extracts from both uninfected and infected HeLa cells supported the accurate initiation of transcription in vitro with almost the same efficiency. In addition, the cell free extract directed an artificial initiation of transcription in vitro by recognizing a TATA box located 140 bp upstream from the cap sites.

摘要

对2型腺病毒（Ad2）的IVa2基因进行了体外转录起始研究，该基因在感染后的中期表达。该基因在帽位点上游约30 bp处不包含TATA框。通过对体外延伸产物进行大小分析和单链核酸酶保护图谱分析，得出结论：体外起始在两个体内帽位点精确发生，效率较低但可检测到。通过使用不含主要晚期启动子的克隆IVa2 DNA，表明IVa2基因体外起始效率低可能不是由于两个基因位置接近，而是可能反映了启动子本身的效率。未感染和感染的HeLa细胞的无细胞提取物以几乎相同的效率支持体外转录的精确起始。此外，无细胞提取物通过识别位于帽位点上游140 bp处的TATA框，在体外指导人工转录起始。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f2f/326990/86d2e95bb6af/nar00391-0050-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f2f/326990/948d50b9f542/nar00391-0046-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f2f/326990/e367e4ae23ff/nar00391-0047-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f2f/326990/ed199149e906/nar00391-0049-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f2f/326990/86d2e95bb6af/nar00391-0050-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f2f/326990/948d50b9f542/nar00391-0046-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f2f/326990/e367e4ae23ff/nar00391-0047-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f2f/326990/ed199149e906/nar00391-0049-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f2f/326990/86d2e95bb6af/nar00391-0050-a.jpg

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本文引用的文献

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Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5216-20. doi: 10.1073/pnas.89.12.5216.
在同源无细胞系统中丝心蛋白基因的忠实转录起始揭示了远上游5'侧翼序列的增强作用。
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Specific in vitro initiation of transcription on the adenovirus type 2 early and late EII transcription units.在腺病毒2型早期和晚期EII转录单元上进行特定的体外转录起始。
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