Heterologous transmembrane and cytoplasmic domains direct functional chimeric influenza virus hemagglutinins into the endocytic pathway.
Heterologous transmembrane and cytoplasmic domains direct functional chimeric influenza virus hemagglutinins into the endocytic pathway.
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DOI：
10.1083/jcb.102.4.1271
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发表时间：
1986-04
期刊：
影响因子：
--
通讯作者：
Gething MJ
中科院分区：
文献类型：
--
作者：
Roth MG;Doyle C;Sambrook J;Gething MJ
通过将编码流感病毒血凝素（HA）的胞外域的DNA序列与编码水疱性口炎病毒的G糖蛋白或单纯疱疹病毒1的gC糖蛋白的跨膜和胞质结构域的DNA融合来产生嵌合基因。用携带重组基因的SV40载体感染的CV-1细胞在其表面表达大量的嵌合蛋白，HAG或HAgC。虽然HAG和HAgC的胞外域在其免疫学性质上与HA不同，但嵌合体显示野生型蛋白的生物学功能特征。HAG和HAgC结合红细胞的效率一样，HA没有，短暂暴露于酸性环境后，诱导红细胞和CV-1细胞膜的融合。然而，HAG和HAgC在细胞表面的行为在几个重要方面与HA不同。观察到HAG和HAgC聚集在包被的凹坑中，而野生型HA被排除在这些结构之外。在氯喹的存在下，它抑制受体从内体的出口，HAG和HAgC积累在细胞内囊泡。相比之下，氯喹对野生型HA的位置没有影响。HAG和HAgC标记在细胞表面表现出温度依赖性收购的电阻细胞外蛋白酶的速率类似于许多细胞表面受体观察到的内化率。HA以至少慢20倍的速率获得对蛋白酶的抗性。我们的结论是，HAG和HAgC有效地路由到内吞途径和HA不是。然而，像HA一样，HAG降解缓慢，这增加了HAG粘附到质膜的可能性。
Chimeric genes were created by fusing DNA sequences encoding the ectodomain of the influenza virus hemagglutinin (HA) to DNA coding for the transmembrane and cytoplasmic domains of either the G glycoprotein of vesicular stomatitis virus or the gC glycoprotein of Herpes simplex virus 1. CV-1 cells infected with SV40 vectors carrying the recombinant genes expressed large amounts of the chimeric proteins, HAG or HAgC on their surfaces. Although the ectodomains of HAG and HAgC differed in their immunological properties from that of HA, the chimeras displayed the biological functions characteristic of the wild-type protein. Both HAG and HAgC bound erythrocytes as efficiently as HA did and, after brief exposure to an acidic environment, induced the fusion of erythrocyte and CV-1 cell membranes. However, the behavior of HAG and HAgC at the cell surface differed from that of HA in several important respects. HAG and HAgC were observed to collect in coated pits whereas wild-type HA was excluded from those structures. In the presence of chloroquine, which inhibits the exit of receptors from endosomes, HAG and HAgC accumulated in intracellular vesicles. By contrast, chloroquine had no effect on the location of wild-type HA. HAG and HAgC labeled at the cell surface exhibited a temperature-dependent acquisition of resistance to extracellular protease at a rate similar to the rates of internalization observed for many cell surface receptors. HA acquired resistance to protease at a rate at least 20- fold slower. We conclude that HAG and HAgC are efficiently routed into the endocytic pathway and HA is not. However, like HA, HAG was degraded slowly, raising the possibility that HAG recycles to the plasma membrane.
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影响因子：
5.4
作者：
FRINK, RJ;EISENBERG, R;WAGNER, EK
通讯作者：
WAGNER, EK
影响因子：
7.8
作者：
HUBBARD, AL;COHN, ZA
通讯作者：
COHN, ZA
影响因子：
7.8
作者：
ALONSOCAPLEN, FV;COMPANS, RW
通讯作者：
COMPANS, RW
影响因子：
64.5
作者：
BASU, SK;GOLDSTEIN, JL;BROWN, MS
通讯作者：
BROWN, MS
DOI：
10.1083/jcb.97.2.329
发表时间：
1983-08
期刊：
The Journal of cell biology
影响因子：
--
作者：
Harding C;Heuser J;Stahl P
通讯作者：
Stahl P