Jelsma T N, Howe J A, Evelegh C M, Cunniff N F, Skiadopoulos M H, Floroff M R, Denman J E, Bayley S T
Department of Biology, McMaster University, Hamilton, Ontario, Canada.
Virology. 1988 Apr;163(2):494-502. doi: 10.1016/0042-6822(88)90290-5.
To help in identifying functional domains within Ad5 E1A proteins, we have constructed a series of mutants that create deletions throughout these products. We have also produced several mis-sense point mutations in the unique 13 S mRNA region. These mutated E1A regions have been tested in plasmid form for their ability to activate transcription of an E3-promoted CAT gene. From the results, a major domain for transactivation has been identified. This begins between residues 138 and 147, ends between residues 188 and 204, and encompasses the unique 13 S region. This domain is sensitive to mis-sense mutations. Transactivation was unaffected by small deletions in the N-terminal half of E1A proteins between residues 4 and 138, but was destroyed when this whole region was deleted. The C-terminal 71 residues may affect transactivation, but the results with the mutant in which this region was deleted were variable. The results obtained with these mutants are discussed in relation to the transactivation obtained by J. W. Lillie et al. [(1987). Cell 50, 1091-1100] with a synthetic peptide similar to the domain described here.
为了有助于鉴定腺病毒5型E1A蛋白中的功能结构域，我们构建了一系列突变体，这些突变体在这些产物中产生了多处缺失。我们还在独特的13S mRNA区域产生了几个错义点突变。这些突变的E1A区域已以质粒形式测试其激活E3启动的CAT基因转录的能力。从结果中，已鉴定出一个主要的反式激活结构域。该结构域起始于第138和147位氨基酸残基之间，终止于第188和204位氨基酸残基之间，并包含独特的13S区域。该结构域对错义突变敏感。E1A蛋白N端一半位于第4和138位氨基酸残基之间的小缺失不影响反式激活，但当整个该区域被缺失时，反式激活被破坏。C端的71个氨基酸残基可能影响反式激活，但该区域被缺失的突变体的结果是可变的。将这些突变体获得的结果与J.W.利利等人[(1987年)。《细胞》50卷，1091 - 1100页]用与本文所述结构域相似的合成肽获得的反式激活结果进行了讨论。
