A 6700 MW membrane protein is encoded by region E3 of adenovirus type 2.
A 6700 MW membrane protein is encoded by region E3 of adenovirus type 2.
复制标题
6700 MW 膜蛋白由 2 型腺病毒 E3 区编码。
DOI：
10.1016/0042-6822(90)90395-8
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发表时间：
1990
期刊：
影响因子：
3.7
通讯作者：
Wold,WS
中科院分区：
文献类型：
--
作者：
Wilson-Rawls,J;Saha,SK;Krajcsi,P;Tollefson,AE;Gooding,LR;Wold,WS
腺病毒2的E3转录单元中，在atg1022和tga1205之间存在一个开放阅读框，可编码分子量为MW 6700 (6.7K)（61个氨基酸）的蛋白。为了确定该蛋白是否表达，我们制备了一种针对6.7K蛋白残基47-61对应的合成肽的抗血清。在10-18%梯度的sds -聚丙烯酰胺凝胶上电泳观察到，该抗血清免疫沉淀了两组蛋白带，7K-8K双链和15K-16K双链或三链。这些条带不是从缺乏6.7K基因的突变体感染的细胞中获得的。大多数（如果不是全部的话）7K-8K和15K-16K条带通过免疫印迹检测到，表明它们是6.7K蛋白的修饰版本。杂交纯化的mRNA在无细胞翻译后只观察到一个8K带，提示这可能是主要的翻译产物。从DNA序列判断，6.7K蛋白具有至少22个残基（残基16-37）的疏水结构域，提示6.7K可能是一个膜蛋白。与此一致的是，在生物化学分离细胞的粗膜中观察到7K-8K和15K-16K条带，而在胞浆和细胞核中没有观察到。突变体产生E3 mrna a和c时，6.7K蛋白产生不足，这表明6.7K是从这些mrna翻译而来的。由于E3-gp19K蛋白也由mrna a和c翻译，因此这些mrna是双链的。6.7K蛋白在Ad5 （Ad2和Ad5是C组腺病毒）中保守性较好，在Ad3 （B组）中保守性较弱。
There is an open reading frame between ATG1022and TGA1205in the E3 transcription unit of adenovirus 2 that could encode a protein of MW 6700 (6.7K) (61 amino acids). To address whether this protein is expressed, we prepared an antiserum against a synthetic peptide corresponding to residues 47–61 in the 6.7K protein. This antiserum immunoprecipitated two series of protein bands, a 7K–8K doublet and a 15K–16K doublet or triplet, as observed by electrophoresis on 10–18% gradient SDS-polyacrylamide gels. These bands were not obtained from cells infected with mutants that lack the 6.7K gene. Most, if not all, of the 7K–8K and 15K–16K bands were detected by immunoblot, indicating that they are modified versions of the 6.7K protein. Only an 8K band was observed after cell-free translation of hybridization-purified mRNA, suggesting that this may be the primary translation product. As judged by DNA sequence, the 6.7K protein has a hydrophobic domain of at least 22 residues (residues 16–37), suggesting that 6.7K may be a membrane protein. Consistent with this, the 7K–8K and 15K–16K bands were observed in the crude membrane but not the cytosol or nuclear fractions of biochemically fractionated cells. The 6.7K protein was underproduced by mutants which under-produce E3 mRNAs a and c, indicating that 6.7K is translated from these mRNAs. Since the E3-gp19K protein is also translated from mRNAs a and c, these mRNAs are bicistronic. The 6.7K protein is well-conserved in Ad5 (Ad2 and Ad5 are group C adenoviruses), and appears to be marginally conserved in Ad3 (group B).