ANALYSIS OF THE PROMOTER AND CIS-ACTING ELEMENTS REGULATING EXPRESSION OF HERPES-SIMPLEX VIRUS TYPE-2 LATENCY-ASSOCIATED TRANSCRIPTS
ANALYSIS OF THE PROMOTER AND CIS-ACTING ELEMENTS REGULATING EXPRESSION OF HERPES-SIMPLEX VIRUS TYPE-2 LATENCY-ASSOCIATED TRANSCRIPTS
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DOI：
10.1128/jvi.69.5.2873-2880.1995
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发表时间：
1995-05-01
影响因子：
5.4
通讯作者：
STRAUS, SE
中科院分区：
文献类型：
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作者：
WANG, K;KRAUSE, PR;STRAUS, SE
在潜伏感染的神经元中，单纯疱疹病毒2型（HSV-2）表达一个丰富的转录物家族，即潜伏相关转录物（LATs）。我们在这里证明，位于HSV-2主要LAT的5'端上游约700 bp的序列在神经元和非神经元细胞的瞬时表达测定中作为非常强的启动子。转录开始于功能性TATA盒下游约27至32 bp处。从-102到+34的近端片段包括基础启动子，并解释了各种细胞系中的组成型转录活性。从-392到-103的远端区域有助于在神经元细胞系中特别强的启动子活性，并且涉及多个顺式作用元件。一个功能性转录激活因子/环腺苷酸（cAMP）反应元件结合蛋白基序位于TATA的上游。通过DNA酶I足迹和甲基化保护分析，我们确定了几个额外的蛋白结合位点上游的激活转录因子/cAMP反应元件结合蛋白基序。富含GC的元件，称为LAT-3，位于碱基-128至-102之间。LAT-3中的2-bp取代显著降低了启动子活性，并在体外消除了蛋白结合能力。凝胶阻滞试验表明，没有竞争的蛋白结合LAT-3的其他GC丰富的元素。LAT-3似乎是一种新的顺式作用元件，可能有助于HSV-2 LAT启动子的神经元反应性。
In latently infected neurons, herpes simplex virus type 2 (HSV-2) expresses one abundant family of transcripts, the latency-associated transcripts (LATs). We demonstrate here that the sequence lying about 700 bp upstream of the 5' end of the HSV-2 major LAT acts as a very strong promoter in transient expression assays in both neuronal and nonneuronal cells. Transcription starts about 27 to 32 bp downstream of a functional TATA box. The proximal fragment from -102 to +34 includes the basal promoter and accounts for constitutive transcriptional activity in various cell lines. The distal region from -392 to -103 contributes to particularly strong promoter activity in neuronal cell lines and involves multiple cis-acting elements. A functional activating transcription factor/cyclic AMP (cAMP) response element binding protein motif lies just upstream of the TATA. By DNase I footprint and methylation protection assays, we identified several additional protein-binding sites upstream of the activating transcription factor/cAMP response element binding protein motif. A GC-rich element, termed LAT-3, was located between bases -128 to -102. A 2-bp substitution in LAT-3 markedly reduced promoter activity and abolished protein-binding ability in vitro. Gel retardation assay showed no competition for protein binding to LAT-3 by other GC-rich elements. LAT-3 appears to be a novel cis-acting element that may contribute to the neuronal responsiveness of the HSV-2 LAT promoter.