ADENOVIRUS 2 CODED EARLY GLYCOPROTEIN

蛋白质通常定位在细胞内的特定位置，并且 因此，调控信息必须存在于蛋白质的结构中 以确保他们到达正确的目的地。我们的主要目标是 是用19 kDa的等离子体作为模型来识别这一信息 早期E3型腺病毒1区(AD2)编码的膜糖蛋白(Gp19K) 或Ad5，并使用遗传方法。Gp19K是一种丰富的蛋白质，其 结构是已知的，活的病毒突变体可以很容易地通过以下方式构建 体外定点诱变。抗血清可用来对抗 纯化的gp19K以及与gp19K相对应的合成肽。 Gp19K有一个17个氨基酸的N端信号序列，两个高甘露糖 寡糖和一个20个氨基酸的疏水跨膜结构域 然后在C末端有一个15个氨基酸的极性区。我们有 分离出的病毒突变体在信号序列中具有测序损伤， 糖基化位点，以及C端疏水和/或极性域。 我们建议从加工和加工的角度来描述这些突变体 运输。现在已经有了一些突变体的数据。在协作环境中 研究表明，这些突变体将被作为病毒抗原类的独特模型进行研究 1 MHC抗原相互作用。其他一些已测序的质粒型突变体如下 可用于构建更多的病毒突变体。我们提出了两部小说和 将细菌遗传学中的标准技术应用于 分离gp19K的随机错义突变。 E3区编码其他几种膜蛋白，它们的结构是 从根本上不同于gp19K。我们建议进行基因研究，类似于 那些带有gp19K的，用来识别膜的结合和运输 这些蛋白质的信号。其中一些基因中的病毒突变， 包括与gp19K的融合，目前有更多的突变体 都很容易建造。针对其中每一种的多肽抗血清 蛋白质是可用的，并且有效的抗血清定向融合蛋白 将生成由表达载体合成的。

Proteins generally are localized at specific sites within the cell, and therefore regulatory information must exist in the structure of the protein to assure that they arrive at the correct destination. Our major objective is to identify this information, using as a model the 19 kDa plasma membrane glycoprotein (gp19K) coded by early region E3 adenovirus 1 (Ad2) or Ad5, and using a genetic approach. Gp19K is an abundant protein whose structure is known, and viable virus mutants can easily be constructed by in vitro site directed mutagenesis. Antisera are available against purified gp19K as well as synthetic peptides corresponding to gp19K. Gp19K has a 17 amino acid N-terminal "signal" sequence, two high-mannose oligosaccharides, and a 20 amino acid hydrophobic transmembrane domain followed by a 15 amino acid polar domain at the C-terminus. We have isolated virus mutants with sequenced lesions in the signal sequence, the glycosylation sites, and the C-terminal hydrophobic and/or polar domains. We propose to characterize these mutants in terms of processing and transport. Data are now available with some mutants. In a collaborative study, the mutants will be studied as a unique model for viral antigenclass 1 MHC antigen interaction. Other plasmid mutants, some sequenced, are available to construct additional virus mutants. We propose two novel and simple methods that apply standard techniques in bacterial genetics to isolate random missense mutations in gp19K. Region E3 encodes several other membrane proteins whose structures are fundamentally different from gp19K. We propose genetic studies, similar to those with gp19K, to identify the membrane-association and transport signals for these proteins. Virus mutants in some of the these genes, including fusions to gp19K, are currently available and additional mutants are easily constructed. Peptide antisera targeted to each of these proteins are available, and potent antisera directed fusion proteins synthesized by expression vectors will be generated.