Mutational analysis of the signal-anchor domain of influenza virus neuraminidase.
Mutational analysis of the signal-anchor domain of influenza virus neuraminidase.
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流感病毒神经氨酸酶信号锚定结构域的突变分析。
DOI：
10.1073/pnas.84.1.1
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发表时间：
1987
影响因子：
11.1
通讯作者：
Nayak,DP
中科院分区：
文献类型：
--
作者：
Sivasubramanian,N;Nayak,DP
流感病毒神经氨酸酶（NA; EC 3.2.1.18）在氨基末端具有信号锚定疏水结构域。为了表征该信号锚定结构域的性质，我们通过寡核苷酸定向诱变在该结构域中引入了单个氨基酸的变化。合成的三种突变NA蛋白在信号锚定结构域的位置11、17或26处含有单个带电氨基酸残基代替疏水性氨基酸残基。当改变的NA蛋白在CV-1细胞中表达时，观察到两种表型：在位置11处精氨酸取代甘氨酸和在位置17处天冬氨酸取代缬氨酸并没有取消信号、锚或运输功能。另一方面，在位置26处用精氨酸取代异亮氨酸阻断了NA蛋白从高尔基复合体向细胞表面的迁移。从这些单点突变构建的双突变体，他们表现出两种表型：一个双突变体（天冬氨酸在位置17和精氨酸在位置26）主要存在于细胞质和其他（精氨酸在位置11和26）主要存在于粗面内质网。这些结果表明，位置11、17和26处的疏水氨基酸是细胞内转运所需的。此外，在粗面内质网或高尔基体的突变蛋白的积累表明存在推定的细胞内运输（或交通）信号中的信号锚定域的NA。
Influenza virus neuraminidase (NA; EC 3.2.1.18) possesses a signal-anchor hydrophobic domain at the amino terminus. To characterize the nature of this signal-anchor domain we have introduced single amino acid changes in this domain by oligonucleotide-directed mutagenesis. Three mutant NA proteins that were synthesized contained a single charged amino acid residue in place of a hydrophobic amino acid residue at position 11, 17, or 26 of the signal-anchor domain. When the altered NA proteins were expressed in CV-1 cells, two phenotypes were observed: substitution of arginine in place of glycine at position 11 and substitution of aspartic acid for valine at position 17 did not abolish the signal, the anchor, or the transport functions. On the other hand, substitution of arginine for isoleucine at position 26 blocked the migration of the NA protein from the Golgi complex to the cell surface. Double mutants were constructed from these single point mutations and they exhibited two phenotypes: one double mutant (aspartic acid at position 17 and arginine at position 26) was present mostly in the cytoplasm and the other (arginine at positions 11 and 26) was present mostly in the rough endoplasmic reticulum. These results indicate that the hydrophobic amino acids at positions 11, 17, and 26 are required for intracellular transport. Furthermore, the accumulation of the mutant proteins in the rough endoplasmic reticulum or the Golgi apparatus suggests the existence of putative intracellular transport (or traffic) signals in the signal-anchor domain of NA.