FUNCTIONAL DISSECTION OF VP16, THE TRANS-ACTIVATOR OF HERPES-SIMPLEX VIRUS IMMEDIATE EARLY GENE-EXPRESSION
FUNCTIONAL DISSECTION OF VP16, THE TRANS-ACTIVATOR OF HERPES-SIMPLEX VIRUS IMMEDIATE EARLY GENE-EXPRESSION
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DOI：
10.1101/gad.2.6.718
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发表时间：
1988-06-01
影响因子：
10.5
通讯作者：
MCKNIGHT, SL
中科院分区：
文献类型：
--
作者：
TRIEZENBERG, SJ;KINGSBURY, RC;MCKNIGHT, SL
1 型单纯疱疹病毒的成熟病毒粒子含有一种激活因子，可启动五个病毒编码的立即早期 (IE) 基因的转录。该激活剂由称为 VP16 的 65 kD 多肽指定。 VP16 的作用是通过位于每个 IE 基因附近区域的顺式调控元件介导的。虽然 VP16 通常通过感染病毒粒子引入细胞，但也可以通过用编码 VP16 的质粒与 IE 顺式调控序列驱动的报告基因共转染细胞来观察其反式激活功能。我们已经使用这样的测定来检查 VP16 突变形式的功能。我们的结果初步鉴定了 VP16 的两个结构域，这两个结构域对其在诱导 IE 基因表达中的作用至关重要。其中一个结构域位于 VP16 的羧基端 78 个氨基酸内，其特征在于其酸性。另一个结构域位于蛋白质的更氨基末端区域，似乎可以调整 VP16 对 IE 基因的特异性。根据本文和随附论文中提出的结果，我们预测 VP16 通过蛋白质：蛋白质而不是蛋白质：DNA 相互作用实现 IE 基因特异性。
Mature virions of herpes simplex virus type 1 contain an activating factor that primes transcription from the five virally encoded immediate early (IE) genes. This activator is specified by a 65-kD polypeptide termed VP16. The action of VP16 is mediated through cis-regulatory elements located in regions adjacent to each IE gene. Although VP16 is normally introduced into cells by infecting virions, its trans-activating function can also be observed by cotransfecting cells with a plasmid that encodes VP16 along with a reporter gene driven by IE cis-regulatory sequences. We have used such an assay to examine the function of mutant forms of VP16. Our results provide tentative identification of two domains of VP16 that are crucial to its role in the induction of IE gene expression. One domain is located within the carboxy-terminal 78 amino acids of VP16 and is characterized by its acidity. Another domain, located in a more amino-terminal region of the protein, appears to tailor the specificity of VP16 for IE genes. According to the results presented in this and the accompanying paper, we predict that VP16 achieves IE gene specificity via protein:protein, rather than protein:DNA, interaction.