Identification of eIF2Bgamma and eIF2gamma as cofactors of hepatitis C virus internal ribosome entry site-mediated translation using a functional genomics approach.
Identification of eIF2Bgamma and eIF2gamma as cofactors of hepatitis C virus internal ribosome entry site-mediated translation using a functional genomics approach.
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使用功能基因组学方法鉴定 eIF2Bgamma 和 eIF2gamma 作为丙型肝炎病毒内部核糖体进入位点介导翻译的辅助因子。
DOI：
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发表时间：
2000
影响因子：
11.1
通讯作者：
F. Wong
中科院分区：
文献类型：
--
作者：
M. Kruger;C. Beger;Q. Li;P. J. Welch;R. Tritz;M. Leavitt;J. Barber;F. Wong
丙型肝炎病毒（HCV）的5'-非翻译区高度保守，折叠成复杂的二级结构，并作为内部核糖体进入位点（IRES）启动HCV蛋白的翻译。我们开发了一个基于随机发夹核酶基因库的选择系统，以确定参与HCV IRES功能的细胞因子。将具有随机靶识别序列的逆转录病毒载体核糖酶文库导入HeLa细胞，稳定表达编码潮霉素B磷酸转移酶基因和单纯疱疹病毒胸苷激酶基因（HSV-tk）的双链结构体。HCV IRES介导了HSV-tk基因的翻译。表达抑制HCV ires介导的HSV-tk翻译的核酶的细胞是通过它们对更昔洛韦和红霉素b的抗性来选择的。这两种核酶可重复地赋予更昔洛韦抗性表型，并被证明可以抑制ires介导的HCV核心蛋白的翻译，但不抑制帽依赖蛋白的翻译或细胞生长。这些核酶的功能靶标分别鉴定为人真核起始因子2B （eIF2Bgamma）和2 （eIF2gamma）的γ亚基。eIF2Bgamma和eIF2gamma参与HCV ires介导的翻译，通过针对这些基因mrna中其他位点的核酶进一步验证。除了鉴定细胞IRES辅助因子外，从该细胞选择系统获得的核酶可直接用于特异性抑制HCV病毒转译，从而促进HCV感染的新抗病毒策略的开发。
The 5'-untranslated region of hepatitis C virus (HCV) is highly conserved, folds into a complex secondary structure, and functions as an internal ribosome entry site (IRES) to initiate translation of HCV proteins. We have developed a selection system based on a randomized hairpin ribozyme gene library to identify cellular factors involved in HCV IRES function. A retroviral vector ribozyme library with randomized target recognition sequences was introduced into HeLa cells, stably expressing a bicistronic construct encoding the hygromycin B phosphotransferase gene and the herpes simplex virus thymidine kinase gene (HSV-tk). Translation of the HSV-tk gene was mediated by the HCV IRES. Cells expressing ribozymes that inhibit HCV IRES-mediated translation of HSV-tk were selected via their resistance to both ganciclovir and hygromycin B. Two ribozymes reproducibly conferred the ganciclovir-resistant phenotype and were shown to inhibit IRES-mediated translation of HCV core protein but did not inhibit cap-dependent protein translation or cell growth. The functional targets of these ribozymes were identified as the gamma subunits of human eukaryotic initiation factors 2B (eIF2Bgamma) and 2 (eIF2gamma), respectively. The involvement of eIF2Bgamma and eIF2gamma in HCV IRES-mediated translation was further validated by ribozymes directed against additional sites within the mRNAs of these genes. In addition to leading to the identification of cellular IRES cofactors, ribozymes obtained from this cellular selection system could be directly used to specifically inhibit HCV viral translation, thereby facilitating the development of new antiviral strategies for HCV infection.
影响因子：
10.5
作者：
Pavitt, GD;Ramaiah, KVA;Hinnebusch, AG
通讯作者：
Hinnebusch, AG
锤头核酶对丙型肝炎病毒 RNA 进行细胞内切割并抑制病毒蛋白翻译。
DOI：
10.1172/jci119097
发表时间：
1996
期刊：
The Journal of clinical investigation.
影响因子：
--
作者：
Sakamoto,N;Wu,CH;Wu,GY
通讯作者：
Wu,GY
DOI：
10.1073/pnas.80.9.2767
发表时间：
1983-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
CHENG, YC;HUANG, ES;GRILL, SP
通讯作者：
GRILL, SP