Kim Dool-Bboon, Zabierowski Susan, DeLuca Neal A
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA.
J Virol. 2002 Feb;76(4):1548-58. doi: 10.1128/jvi.76.4.1548-1558.2002.
The start site regions of late genes of herpes simplex virus type 1 are similar to the eukaryotic initiator sequence (Inr), have been shown to affect the levels of expression, and may also play a role in transcription activation by the viral activator ICP4. A series of linker-scanning mutations spanning the start site of transcription and several downstream mutations in the true late gC promoter were analyzed in reconstituted in vitro transcription reactions with and without ICP4, as well as in the context of the viral genome during infection. The nucleotide contacts previously found to be important for Inr function were also found to be important for optimal induction by ICP4. While the Inr had a substantial effect on the accumulation of gC RNA during infection, no other sequence downstream of the TATA box to +124 had a significant effect on levels of expression during infection. Therefore, these studies suggest that TATA box and the Inr are the only cis-acting elements required to achieve optimal expression of gC, and that the high levels of late-gene transcription may be largely due to the induction by ICP4, functioning through the Inr element.
单纯疱疹病毒1型晚期基因的起始位点区域与真核生物起始子序列(Inr)相似,已被证明会影响表达水平,并且可能在病毒激活因子ICP4介导的转录激活过程中发挥作用。在有无ICP4参与的体外重组转录反应中,以及在感染期间病毒基因组的背景下,分析了一系列跨越转录起始位点的接头扫描突变以及真正晚期gC启动子中的几个下游突变。先前发现对Inr功能重要的核苷酸接触位点,对于ICP4的最佳诱导作用也很重要。虽然Inr在感染期间对gC RNA的积累有显著影响,但TATA框下游至+124的其他序列在感染期间对表达水平没有显著影响。因此,这些研究表明,TATA框和Inr是实现gC最佳表达所需的唯一顺式作用元件,并且晚期基因的高转录水平可能很大程度上归因于ICP4通过Inr元件介导的诱导作用。