Department of Cellular and Molecular Medicine and Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, La Jolla, California 92093-0712, USA.
J Virol. 2010 May;84(10):5158-70. doi: 10.1128/JVI.00090-10. Epub 2010 Mar 3.
It has previously been demonstrated that, during human cytomegalovirus infection, the viral IE2 86 and IE2 40 proteins are both important for the expression of an early-late viral protein, UL84. Here, we show that expression of the UL84 protein is enhanced upon cotransfection with either IE2 86 or IE2 40, although IE2 40 appears to play a more important role. The UL84 protein levels are tightly linked to the amount of IE2 40 present, but this does not appear to be true for IE2 86. RNA remains constant for all corresponding proteins, indicating posttranscriptional regulation of UL84. The first 105 amino acids of UL84 are necessary and sufficient for this phenotype, and this region is also required for an interaction with IE2 86 and IE2 40. Treatment with proteasome inhibitors shows that UL84 exhibits some proteasome-dependent degradation, and UL84 is not protected against this degradation when coexpressed with IE2 86 or IE2 40. UL84 also exhibits an inhibitory effect on IE2 86 and IE2 40 protein levels in these cotransfection assays. Further, we show that the amino acid sequence of UL84 is important for the enhancement governed by IE2 40. These results indicate that IE2 86, IE2 40, and UL84 serve to regulate protein expression in a posttranscriptional fashion and that this regulation is independent of other viral proteins.
先前的研究表明,在人类巨细胞病毒感染过程中,病毒 IE2 86 和 IE2 40 蛋白对于早期晚期病毒蛋白 UL84 的表达都是重要的。在这里,我们表明,在共转染 IE2 86 或 IE2 40 时,UL84 蛋白的表达会增强,尽管 IE2 40 似乎发挥更重要的作用。UL84 蛋白水平与存在的 IE2 40 量紧密相关,但对于 IE2 86 则并非如此。所有相应蛋白的 RNA 保持不变,表明 UL84 的转录后调控。UL84 的前 105 个氨基酸是这种表型所必需和充分的,该区域也需要与 IE2 86 和 IE2 40 相互作用。用蛋白酶体抑制剂处理表明 UL84 表现出一些蛋白酶体依赖性降解,并且当与 IE2 86 或 IE2 40 共表达时,UL84 不受这种降解的保护。UL84 在这些共转染实验中还对 IE2 86 和 IE2 40 蛋白水平表现出抑制作用。此外,我们表明 UL84 的氨基酸序列对于由 IE2 40 控制的增强作用很重要。这些结果表明,IE2 86、IE2 40 和 UL84 在后转录水平上调节蛋白表达,并且这种调节独立于其他病毒蛋白。