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马疱疹病毒1型立即早期基因产物截短版本的核定位与转录激活活性

Nuclear localization and transcriptional activation activities of truncated versions of the immediate-early gene product of equine herpesvirus 1.

作者信息

Smith R H, Holden V R, O'Callaghan D J

机构信息

Department of Microbiology and Immunology, Louisiana State University Medical Center, Shreveport 71130-3932, USA.

出版信息

J Virol. 1995 Jun;69(6):3857-62. doi: 10.1128/JVI.69.6.3857-3862.1995.

Abstract

The equine herpesvirus 1 (EHV-1) immediate-early (IE) gene product encodes a nuclear regulatory protein capable of negatively autoregulating its own promoter, transactivating representative EHV-1 early promoters, and acting in a concerted fashion with accessory EHV-1 regulatory factors to transactivate EHV-1 late promoters. To identify IE amino acid sequences involved in nuclear localization and to examine the contribution of C-terminal portions of the IE polypeptide to transactivation, vectors that express various carboxyterminally truncated IE polypeptides were constructed. It is demonstrated that amino acids 963 through 970 of the 1,487-amino-acid IE protein are required for efficient localization of the truncated IE polypeptides to the nuclei of transfected cells. In addition, it is demonstrated that the first 970 amino acids of the IE gene product are sufficient to transactivate the EHV-1 thymidine kinase promoter to significant levels (i.e., approximately 40% of the level of wild-type activation).

摘要

马疱疹病毒1型(EHV-1)立即早期(IE)基因产物编码一种核调节蛋白,该蛋白能够对自身启动子进行负向自调节、反式激活代表性的EHV-1早期启动子,并与EHV-1辅助调节因子协同作用以反式激活EHV-1晚期启动子。为了鉴定参与核定位的IE氨基酸序列,并研究IE多肽C末端部分对反式激活的贡献,构建了表达各种羧基末端截短的IE多肽的载体。结果表明,1487个氨基酸的IE蛋白中963至970位氨基酸是截短的IE多肽有效定位到转染细胞核所必需的。此外,结果表明IE基因产物的前970个氨基酸足以将EHV-1胸苷激酶启动子反式激活到显著水平(即,约为野生型激活水平的40%)。

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