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酵母Dbp2p“DEAD-box”蛋白表达的自动调节由保守的Dbp2内含子中的序列介导。

Autoregulation of expression of the yeast Dbp2p 'DEAD-box' protein is mediated by sequences in the conserved DBP2 intron.

作者信息

Barta I, Iggo R

机构信息

Swiss Institute for Experimental Cancer Research (ISREC), Epalinges.

出版信息

EMBO J. 1995 Aug 1;14(15):3800-8. doi: 10.1002/j.1460-2075.1995.tb00049.x.

Abstract

The human p68, Saccharomyces cerevisiae DBP2 and Schizosaccharomyces pombe dbp2 genes are closely related members of the 'DEAD-box' RNA helicase superfamily. All three genes contain an intron at a conserved site in RNA helicase motif V. The S.cerevisiae intron is unusual both for its position near the 3'-end of the open reading frame and for its size, 1001 nucleotides. We show here that precise deletion of the intron has no effect on cell viability but leads to an increase in Dbp2p protein expression. Inefficient splicing due to the size of the intron can not account for this difference because the intron is efficiently spliced in Dbp2p-deficient cells. Instead, there is a reciprocal relationship between the amount of Dbp2p in the cell and the efficiency with which DBP2 intron-containing genes are expressed. Inactive Dbp2p mutants are efficiently expressed from DBP2 intron-containing plasmids, and fragments of the DBP2 intron confer Dbp2p-responsiveness on heterologous reporter introns. This suggest that there is an intron-mediated negative feedback loop regulating DBP2 expression, and provides a possible explanation for the retention of such an unusual intron in S.cerevisiae.

摘要

人类p68、酿酒酵母DBP2和粟酒裂殖酵母dbp2基因是“DEAD-box”RNA解旋酶超家族中密切相关的成员。这三个基因在RNA解旋酶基序V的保守位点都含有一个内含子。酿酒酵母的内含子不同寻常,这不仅体现在其位于开放阅读框3'端附近的位置,还体现在其大小上,为1001个核苷酸。我们在此表明,精确缺失该内含子对细胞活力没有影响,但会导致Dbp2p蛋白表达增加。由于内含子大小导致的低效剪接并不能解释这种差异,因为该内含子在缺乏Dbp2p的细胞中能有效剪接。相反,细胞中Dbp2p的量与含DBP2内含子基因的表达效率之间存在相互关系。无活性的Dbp2p突变体从含DBP2内含子的质粒中能有效表达,并且DBP2内含子的片段赋予异源报告内含子Dbp2p反应性。这表明存在一个内含子介导的负反馈环调节DBP2的表达,并为酿酒酵母中保留这样一个不寻常的内含子提供了一种可能的解释。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7208/394454/6fd325a30543/emboj00039-0215-a.jpg

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