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小鼠c-fos基因第一个内含子内过早转录终止的序列要求。

Sequence requirements for premature transcription arrest within the first intron of the mouse c-fos gene.

作者信息

Mechti N, Piechaczyk M, Blanchard J M, Jeanteur P, Lebleu B

机构信息

Laboratoire de Biochimie des Protéines, URA CNRS 1191, Génétique Moléculaire, Université des Sciences et Techniques du Languedoc, Montpellier, France.

出版信息

Mol Cell Biol. 1991 May;11(5):2832-41. doi: 10.1128/mcb.11.5.2832-2841.1991.

Abstract

A strong block to the elongation of nascent RNA transcripts by RNA polymerase II occurs in the 5' part of the mammalian c-fos proto-oncogene. In addition to the control of initiation, this mechanism contributes to transcriptional regulation of the gene. In vitro transcription experiments using nuclear extracts and purified transcription templates allowed us to map a unique arrest site within the mouse first intron 385 nucleotides downstream from the promoter. This position is in keeping with that estimated from nuclear run-on assays performed with short DNA probes and thus suggests that it corresponds to the actual block in vivo. Moreover, we have shown that neither the c-fos promoter nor upstream sequences are absolute requirements for an efficient transcription arrest both in vivo and in vitro. Finally, we have characterized a 103-nucleotide-long intron 1 motif comprising the arrest site and sufficient for obtaining the block in a cell-free transcription assay.

摘要

RNA聚合酶II对新生RNA转录本延伸的强烈阻滞发生在哺乳动物原癌基因c-fos的5'端区域。除了起始控制外,这种机制也有助于该基因的转录调控。利用核提取物和纯化的转录模板进行的体外转录实验,使我们能够在小鼠第一个内含子中定位到一个独特的阻滞位点,该位点位于启动子下游385个核苷酸处。这个位置与用短DNA探针进行的核延伸实验所估计的位置一致,因此表明它对应于体内实际的阻滞位点。此外,我们已经表明,无论是c-fos启动子还是上游序列,对于体内和体外的有效转录阻滞都不是绝对必需的。最后,我们鉴定了一个103个核苷酸长的内含子1基序,它包含阻滞位点,并且足以在无细胞转录实验中产生阻滞作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2587/360067/ab518552da1d/molcellb00139-0510-a.jpg

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