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两种转录方向相反的盘基网柄菌基因对的表征以及对位于它们之间具有基础启动子元件特征的富含G序列元件的鉴定。

Characterization of two divergently transcribed Dictyostelium gene pairs and identification of G-rich sequence element lying between them with the characteristics of a basal promoter element.

作者信息

Driscoll D M, Pears C J, Williams J G

机构信息

Imperial Cancer Research Fund, Clare Hall Laboratories, Potters Bar, Hertsfordshire, England.

出版信息

Dev Genet. 1988;9(4-5):455-68. doi: 10.1002/dvg.1020090423.

Abstract

The cysteine proteinase 1 (CP1) and cysteine proteinase 2 (CP2) genes of Dictyostelium discoideum encode coordinately expressed mRNA sequences that are inducible by extracellular cAMP. Both genes form part of divergently transcribed gene pairs. The gene proximal to CP1 is coordinately regulated and encodes a protein containing several potential zinc binding domains of the kind found in DNA binding proteins. The gene proximal to CP2 is a constitutively transcribed gene of unknown function. There are multiple, short, G-rich sequence elements between both gene pairs, and deletion of the pair of elements 200 nucleotides upstream from the CP2 gene abolishes cAMP-inducibility. A synthetic oligonucleotide, containing two copies of the G-rich element from the CP1 gene, will reconstitute cAMP-inducibility in the deletion mutant of the CP2 gene. This shows that the elements in the two genes are functionally homologous. Efficient induction requires at least two copies of the CP1 element, but their relative orientation is unimportant. Two copies in an inverted orientation are, however, inactive when moved upstream of their normal position and are incapable of conferring cAMP-inducibility on a heterologous gene. These observations suggest that these sequences are either essential promoter elements, not themselves interacting with the inducer, or that their interaction with a separate class of control sequences is necessary for inducible expression.

摘要

盘基网柄菌的半胱氨酸蛋白酶1(CP1)和半胱氨酸蛋白酶2(CP2)基因编码由细胞外cAMP诱导的协同表达的mRNA序列。这两个基因都是反向转录基因对的一部分。CP1近端的基因受到协同调控,编码一种含有几种潜在锌结合结构域的蛋白质,这种结构域存在于DNA结合蛋白中。CP2近端的基因是一个功能未知的组成型转录基因。两个基因对之间有多个短的富含G的序列元件,删除CP2基因上游200个核苷酸处的元件对会消除cAMP诱导性。一个含有来自CP1基因的两个富含G元件拷贝的合成寡核苷酸,将在CP2基因的缺失突变体中重建cAMP诱导性。这表明两个基因中的元件在功能上是同源的。有效的诱导至少需要两个CP1元件拷贝,但它们的相对方向并不重要。然而,两个反向的拷贝在移至其正常位置上游时无活性,并且不能赋予异源基因cAMP诱导性。这些观察结果表明,这些序列要么是必需的启动子元件,本身不与诱导剂相互作用,要么它们与另一类控制序列的相互作用对于诱导表达是必需的。

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