Mutations that alter both localization and production of a yeast nuclear protein.
Mutations that alter both localization and production of a yeast nuclear protein.
复制标题
改变酵母核蛋白的定位和产生的突变。
DOI：
10.1101/gad.2.6.707
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发表时间：
1988
影响因子：
10.5
通讯作者：
Sadler,I
中科院分区：
文献类型：
--
作者：
Silver,PA;Chiang,A;Sadler,I
酵母GAL4基因产物的前74个氨基酸足以将GAL4-β-半乳糖苷酶嵌合蛋白定位到酵母核。嵌合蛋白缺少前74个GAL4氨基酸，但包含GAL4的几乎所有其余部分，不定位于细胞核，并且表达水平高于核对应蛋白。在此基础上，分离和测序了GAL4内的点突变，这些点突变减少了核定位，增加了正常核GAL4-β-半乳糖苷酶融合蛋白的产量。检测了这些突变对完整的GAL4蛋白定位和表达的影响。突变的蛋白质被排除在细胞核之外的程度各不相同，但所有的突变都会导致蛋白质的过度生产。GAL4的六个半胱氨酸残基中的两个点突变可以通过完整的GAL4取消基因激活；然而，附近残基的突变对GAL4依赖的基因激活没有影响。
The first 74 amino acids of the yeast GAL4 gene product are sufficient to localize a GAL4-beta-galactosidase chimeric protein to the yeast nucleus. Chimeric proteins missing the first 74 GAL4 amino acids, but containing almost all of the rest of GAL4, are not localized to the nucleus and are expressed at higher levels than their nuclear counterparts. On this basis, point mutations within GAL4, which reduce nuclear localization and increase production of a normally nuclear GAL4-beta-galactosidase fusion protein, were isolated and sequenced. The effect of these mutations on the localization and expression of the intact GAL4 protein was examined. The degree to which the mutant proteins are excluded from the nucleus varies, but all mutations cause overproduction of the protein. Point mutations altering two of the six cysteine residues of the GAL4 putative 'zinc finger' abolish gene activation by intact GAL4; however, mutations in nearby residues have no effect on GAL4-dependent gene activation.