Distal protein sequences can affect the function of a nuclear localization signal.
Distal protein sequences can affect the function of a nuclear localization signal.
复制标题
远端蛋白质序列可以影响核定位信号的功能。
DOI：
10.1128/mcb.12.3.1330-1339.1992
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发表时间：
1992
影响因子：
5.3
通讯作者：
Knipe,DM
中科院分区：
文献类型：
--
作者：
Gao,M;Knipe,DM
由单纯疱疹病毒编码的主要DNA结合蛋白或感染细胞蛋白8（ICP8）可以独立于其他病毒蛋白定位于细胞核。为了确定ICP8内的核定位信号，我们对克隆的ICP8基因进行了几种形式的诱变。ICP8基因的缺失分析表明，ICP8的几个部分参与其核定位。为了确定这些区域是否是独立的定位信号，我们将ICP8基因的各个部分引入一系列盒质粒中，所述盒质粒允许表达含有丙酮酸激酶的融合蛋白，所述丙酮酸激酶通常是与ICP8的各个部分融合的细胞质蛋白。这些结果表明，羧基末端28个残基是ICP8能够靶向蛋白激酶进入细胞核的唯一部分。然而，将ICP8的某些额外区域包含到融合蛋白中导致核定位的抑制。因此，ICP 8的羧基末端28个残基可以独立地作为核定位信号，但是蛋白质的其余部分中的某些构象约束或折叠或组装要求可以影响蛋白质的核定位。我们的结果表明，远离核定位信号的序列可以影响其功能。已分离出一组融合载体，其应普遍用于在任何阅读框架中进行5 '或3'融合以快速定位信号。
The major DNA-binding protein, or infected-cell protein 8 (ICP8), encoded by herpes simplex virus can localize to the cell nucleus independently of other viral proteins. To define the nuclear localization signals within ICP8, we performed several forms of mutagenesis on the cloned ICP8 gene. Deletion analysis of the ICP8 gene showed that several portions of ICP8 are involved in its nuclear localization. To determine whether these regions were independent localization signals, we introduced various portions of the ICP8 gene into a series of cassette plasmids which allowed expression of fusion proteins containing pyruvate kinase, normally a cytoplasmic protein, fused to various portions of ICP8. These results showed that the carboxyl-terminal 28 residues are the only portion of ICP8 capable of targeting protein kinase into the nucleus. However, inclusion of certain additional regions of ICP8 into the fusion protein led to an inhibition of nuclear localization. Therefore, the carboxyl-terminal 28 residues of ICP8 can act independently as a nuclear localization signal, but certain conformational constraints or folding or assembly requirements in the remainder of the protein can affect the nuclear localization of the protein. Our results demonstrate that sequences distant from a nuclear localization signal can affect its ability to function. A set of fusion vectors has been isolated which should be of general use for making 5'or 3'fusions in any reading frame to rapidly map localization signals.