Hepatitis B virus HBx protein localized to the nucleus restores HBx-deficient virus replication in HepG2 cells and in vivo in hydrodynamically-injected mice.
Hepatitis B virus HBx protein localized to the nucleus restores HBx-deficient virus replication in HepG2 cells and in vivo in hydrodynamically-injected mice.
复制标题
丙型肝炎病毒HBX蛋白局部局部在核中恢复HBX缺陷病毒在HEPG2细胞中的复制和体内注入流体动力学小鼠的体内。
DOI：
10.1016/j.virol.2009.05.001
复制
发表时间：
2009-07-20
期刊：
影响因子：
3.7
通讯作者：
Slagle, Betty L.
中科院分区：
文献类型：
--
作者：
Keasler, Victor V.;Hodgson, Amanda J.;Madden, Charles R.;Slagle, Betty L.
确定乙型肝炎病毒 (HBV) 复制中调节 HBx 蛋白的要求是一个重要目标。基于质粒的 HBV 复制测定用于评估 HBx 亚细胞定位是否影响其促进病毒复制的能力，通过病毒衣壳相关 DNA 的实时 PCR 定量来测量。通过核定位信号靶向细胞核的 HBx (NLS-HBx) 能够恢复 HBx 缺陷的 HBV 复制，而含有核输出信号的 HBx (NES-HBx) 则不能。 NLS-HBx 和 NES-HBx 均以相似的水平表达（通过免疫沉淀和蛋白质印迹），并且通过使用与 GFP 融合的 HBx、NLS-HBx 和 NES-HBx 蛋白的反卷积显微镜来确认信号序列标记蛋白的正确定位。重要的是，这些发现通过小鼠体内的流体动力学注射得到了体内证实。我们的结果表明，在这些 HBV 复制测定中，HBx 的至少一项功能需要其定位于细胞核。
Identifying the requirements for the regulatory HBx protein in hepatitis B virus (HBV) replication is an important goal. A plasmid-based HBV replication assay was used to evaluate whether HBx subcellular localization influences its ability to promote virus replication, as measured by real time PCR quantitation of viral capsid-associated DNA. HBx targeted to the nucleus by a nuclear localization signal (NLS-HBx) was able to restore HBx-deficient HBV replication, while HBx containing a nuclear export signal (NES-HBx) was not. Both NLS-HBx and NES-HBx were expressed at similar levels (by immunoprecipitation and Western blotting), and proper localization of the signal sequence-tagged proteins was confirmed by deconvolution microscopy using HBx, NLS-HBx, and NES-HBx proteins fused to GFP. Importantly, these findings were confirmed in vivo by hydrodynamic injection into mice. Our results demonstrate that in these HBV replication assays, at least one function of HBx requires its localization to the nucleus.
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影响因子：
5.4
作者：
Becker, SA;Lee, TH;Slagle, BL
通讯作者：
Slagle, BL
DOI：
10.1073/pnas.90.17.8078
发表时间：
1993-09-01
影响因子：
11.1
作者：
CROSS, JC;WEN, P;RUTTER, WJ
通讯作者：
RUTTER, WJ
影响因子：
3.8
作者：
Henkler, F;Hoare, J;King, IA
通讯作者：
King, IA
影响因子：
12.7
作者：
Hoare, J;Henkler, F;McGarvey, MJ
通讯作者：
McGarvey, MJ
影响因子：
5.4
作者：
Keasler, Victor V.;Hodgson, Amanda J.;Slagle, Betty L.
通讯作者：
Slagle, Betty L.