Amplification of recombinant adenoviral transgene products occurs by inhibition of histone deacetylase
Amplification of recombinant adenoviral transgene products occurs by inhibition of histone deacetylase
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DOI：
10.1006/viro.1997.8538
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发表时间：
1997-05-12
期刊：
影响因子：
3.7
通讯作者：
Garver, RI
中科院分区：
文献类型：
--
作者：
Dion, LD;Goldsmith, KT;Garver, RI
正丁酸盐（丁酸盐）已显示在用El缺陷型腺病毒感染的细胞中扩增转基因表达。进行本研究是为了更好地定义丁酸在腺病毒基因表达背景下的作用，并试图阐明丁酸介导转基因扩增的机制。发现丁酸盐在0.5-5 mM的浓度范围内扩增病毒转基因表达，并且扩增需要暴露12-24小时以获得最大效果。代表性的病毒蛋白质的Western印迹分析表明，丁酸处理扩增DNA结合蛋白，但不是纤维蛋白。瞬时腺病毒复制系统表明丁酸盐对El缺陷型腺病毒的复制具有适度的抑制作用。使用组蛋白去乙酰化酶的特异性抑制剂，抑制素A（TSA），再现了用丁酸酯实现的病毒转基因产物的扩增。相反，在已知具有TSA抗性组蛋白脱乙酰酶的细胞系中，TSA处理不能扩增腺病毒转基因表达。丁酸扩增病毒转基因的稳态基因表达，但对DNA结合蛋白或纤维稳态基因表达没有可检测的影响。核溢流实验表明丁酸盐和TSA均引起病毒转基因转录的增加。结论是组蛋白去乙酰化酶抑制剂在转录水平上放大腺病毒转基因表达。(C)北京：科学出版社.
n-Butyrate (butyrate) has been shown to amplify transgene expression in cells infected with El-defective adenoviruses. The present studies were undertaken in order to better define the actions of butyrate in the context of adenovirus gene expression, and to attempt to elucidate the mechanism by which butyrate mediates the transgene amplification. It was found that butyrate amplified viral transgene expression over a concentration range of 0.5-5 mM, and that the amplification required an exposure of 12-24 hr for maximal effect. Western blot analysis of representative viral proteins showed that butyrate treatment amplified DNA-binding protein, but not fiber protein. A transient adenoviral replication system suggested that butyrate had a modest inhibitory effect on replication of the El-defective adenovirus. Use of a specific inhibitor of histone deacetylase, trichostatin A (TSA), reproduced the amplification of the viral transgene product achieved with the butyrate. In contrast, adenoviral transgene expression could not be amplified by TSA treatment in a cell line known to have a TSA-resistant histone deacetylase. Butyrate amplified steady-state gene expression of the viral transgene, but had no detectable effects on either DNA-binding protein or fiber steady-state gene expression. Nuclear run-off experiments showed that both butyrate and TSA caused an increase in the viral transgene transcription. It was concluded that inhibitors of histone deacetylase amplify adenoviral transgene expression at the transcriptional level. (C) 1997 Academic Press.