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人类c-myc基因中的转录延伸受非洲爪蟾卵母细胞中整体转录起始水平的调控。

Transcription elongation in the human c-myc gene is governed by overall transcription initiation levels in Xenopus oocytes.

作者信息

Spencer C A, Kilvert M A

机构信息

Department of Biochemistry, University of Alberta, Edmonton, Canada.

出版信息

Mol Cell Biol. 1993 Feb;13(2):1296-305. doi: 10.1128/mcb.13.2.1296-1305.1993.

Abstract

Both transcription initiation and transcription elongation contribute to the regulation of steady-state c-myc RNA levels. We have used the Xenopus oocyte transcription assay to study premature transcription termination which occurs in the first exon and intron of the human c-myc gene. Previous studies showed that after injection into Xenopus oocytes transcription from the c-myc P1 promoter resulted in read-through transcripts whereas transcription from the stronger P2 promoter resulted in a combination of prematurely terminated and read-through transcripts. We now demonstrate that this promoter-specific processivity results from the overall amount of RNA polymerase II transcription occurring from either promoter. Parameters that reduce the amount of transcription from P1 or P2, such as decreased concentration of template injected or decreased incubation time, result in a reduction in the ratio of terminated to read-through c-myc transcripts. Conversely, when transcription levels are increased by higher concentrations of injected template, increased incubation time, or coinjection with competing template, the ratio of terminated to read-through transcripts increases. We hypothesize that an RNA polymerase II processivity function is depleted above a threshold level of transcription initiation, resulting in high levels of premature transcription termination. These findings account for the promoter-specific effects on transcription elongation previously seen in this assay system and suggest a mechanism whereby limiting transcription elongation factors may contribute to transcription regulation in other eukaryotic cells.

摘要

转录起始和转录延伸都有助于调节c-myc RNA的稳态水平。我们利用非洲爪蟾卵母细胞转录试验来研究人类c-myc基因第一外显子和内含子中发生的过早转录终止。先前的研究表明,将c-myc P1启动子注射到非洲爪蟾卵母细胞后进行转录会产生通读转录本,而从更强的P2启动子转录则会产生过早终止和通读转录本的组合。我们现在证明,这种启动子特异性的持续性是由任一启动子发生的RNA聚合酶II转录的总量导致的。降低从P1或P2转录量的参数,如降低注射模板的浓度或缩短孵育时间,会导致c-myc转录本中终止与通读的比例降低。相反,当通过更高浓度的注射模板、延长孵育时间或与竞争模板共注射来提高转录水平时,终止与通读转录本的比例会增加。我们推测,RNA聚合酶II的持续性功能在转录起始的阈值水平以上会被耗尽,从而导致高水平的过早转录终止。这些发现解释了此前在该试验系统中观察到的启动子对转录延伸的特异性影响,并提示了一种机制,即有限的转录延伸因子可能在其他真核细胞的转录调控中发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c4a5/359015/4c9a8784d587/molcellb00014-0583-a.jpg

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