REGULATION OF HERPES-SIMPLEX VIRUS-1 GENES - ALPHA-GENE SEQUENCE REQUIREMENTS FOR TRANSIENT INDUCTION OF INDICATOR GENES REGULATED BY BETA-PROMOTERS OR LATE (GAMMA2) PROMOTERS
REGULATION OF HERPES-SIMPLEX VIRUS-1 GENES - ALPHA-GENE SEQUENCE REQUIREMENTS FOR TRANSIENT INDUCTION OF INDICATOR GENES REGULATED BY BETA-PROMOTERS OR LATE (GAMMA2) PROMOTERS
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DOI：
10.1016/0042-6822(86)90117-0
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发表时间：
1986-03-01
期刊：
影响因子：
3.7
通讯作者：
ROIZMAN, B
中科院分区：
文献类型：
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作者：
MAVROMARANAZOS, P;SILVER, S;ROIZMAN, B
该实验室早先报道，由融合到晚期（γ 2）基因的启动子调节结构域的胸苷激酶（TK）基因的结构序列组成的嵌合基因，当驻留在单纯疱疹病毒1基因组中时，被调节为真正的γ 2基因，但不能与β区分。当通过转染引入并稳定整合到宿主基因组的环境中时，银和B。罗伊兹曼Cell. 5，518-528，1985）。我们在此报告，β. -转染到TK-幼仓鼠肾（BHKtk-）中的TK和嵌合γ 2-TK基因被α 4和α O诱导，但不被其它α诱导。基因.具体而言：（i）两种TK基因都是通过与携带完整α 4或完整α O基因的DNA片段共转染诱导的，但不是通过仅携带启动子调节结构域或α 4基因或完整α 22、α 27和α 47基因的结构序列的片段共转染诱导的。(ii)在其3 ″编码序列中携带2700-bp缺失的α 4基因也诱导这两种基因，尽管效率较低。(iii)从用完整或截短的α 4基因转染的细胞的细胞质中回收的与α 4基因同源的RNA，定位于α 4基因转录起始的真正位点。从与TK基因融合的bonafide γ 2基因加帽位点转录与从用γ 2-TK和α 4基因转染的细胞的细胞质中提取的嵌合TK基因同源的RNA。这些结果表明，α 4基因和α O基因各自能够诱导β 4和β O基因的表达。和驻留在细胞基因组环境中的γ 2基因，负责诱导的活性位点位于α 4蛋白的N末端附近，并加强了驻留在宿主细胞环境中的γ 2基因不能用于通过目前可用的测试鉴定γ 2基因调节的真实决定子的结论。
This laboratory reported earlier that chimeric genes consisting of the structural sequences of the thymidine kinase (TK) gene fused to the promoter-regulatory domains of late (.gamma.2) genes were regulated as bonafide .gamma.2 genes when resident in the herpes simplex virus 1 genome but could not be differentiated from .beta. genes when introduced by transfection and stably integrated into the environment of the host genome (S. Silver and B. Roizman, Mol. Cell. Biol. 5, 518-528, 1985). We report here that .beta.-TK and the chimeric .gamma.2-TK gene transfected into TK- baby hamster kidney (BHKtk-) were induced by .alpha.4 and .alpha.O but not by the other .alpha. genes. Specifically: (i) Both TK gnees were induced by cotransfection with DNA fragments carrying an intact .alpha.4 or an intact .alpha.O gene, but not by fragments carrying only the promoter-regulatory domain or the structural sequences of the .alpha.4 gene or intact .alpha.22, .alpha.27, and .alpha.47 genes. (ii) An .alpha.4 gene carrying a 2700-bp deletion in its 3'' coding sequence also induced both genes, although less efficiently. (iii) RNA homologous to the .alpha.4 gene recovered from the cytoplasm of cell transfected with either the intact or truncated .alpha.4 gene mapped to the bonafide site of transcription initiation of the .alpha.4 gene. RNA homologous to the chimeric TK gene extracted from the cytoplasm of cells transfected with both .gamma.2-TK and the .alpha.4 gene was transcribed from the bonafide .gamma.2 gene capping site fused to the TK gene. These results indicate that the .alpha.4 gene and the .alpha.O gene are each capable of inducing the expression of both .beta. and .gamma.2 genes resident in the environment of the cellular genome, that the active site responsible for induction is located near the N terminus of the .alpha.4 protein, and reinforce the conclusion that .gamma.2 genes resident in the environment of the host cell cannot be used to identify the authentic determinants of .gamma.2 gene regulation by currently available tests.