Analysis of prototype foamy virus particle-host cell interaction with autofluorescent retroviral particles.
Analysis of prototype foamy virus particle-host cell interaction with autofluorescent retroviral particles.
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DOI：
10.1186/1742-4690-7-45
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发表时间：
2010-05-17
期刊：
影响因子：
3.3
通讯作者：
Lindemann D
中科院分区：
文献类型：
--
作者：
Stirnnagel K;Lüftenegger D;Stange A;Swiersy A;Müllers E;Reh J;Stanke N;Grosse A;Chiantia S;Keller H;Schwille P;Hanenberg H;Zentgraf H;Lindemann D
泡沫病毒(FV)的复制周期显示出几个独特的特征，这使它们有别于正反转录病毒。首先，像其他B/D型正反转录病毒一样，FV衣壳在中心体预组装，但更类似于Hepadna病毒，FV出芽严格依赖于同源病毒糖蛋白的共表达。其次，FV异常广泛的宿主范围被认为是由于使用了宿主细胞质膜上存在的一种非常常见的进入受体，因为到目前为止，所有体外测试的细胞系都是允许的。为了利用现代荧光显微镜技术来研究FV复制，我们创造了带有各种蛋白质标签的FV Gag蛋白，并评估了它们支持FV复制的各个步骤的能力。在Fv Gag中添加即使是很小的N-末端HA标记也会严重影响Fv颗粒的释放。例如，尽管表面上细胞内的衣壳组装正常，但N-末端的自身荧光蛋白(AFP)融合完全取消了释放。相反，C-末端Gag-tag对颗粒组装、出口和颗粒形态发生的影响很小。与野生型相比，C末端衣壳标记的FV载体颗粒的感染性降低了100倍，但通过野生型Gag的共表达和混合颗粒的组装拯救了感染性。荧光抗体颗粒与靶细胞的特异性结合呈剂量依赖性，依赖于环境，但不与靶细胞提取或合成的脂类结合。对不同来源的靶细胞进行筛选，鉴定出两个细胞系，一个是人红系前体细胞，另一个是斑马鱼细胞系，它们对FV Env介导的FV和HIV载体转导具有抵抗力。我们已经建立了功能性的、自发荧光的泡沫病毒颗粒，作为利用现代荧光成像技术研究FV-宿主细胞相互作用的有价值的新工具。此外，我们首次成功地鉴定了两个对泡沫病毒包膜病毒原型介导的基因转移具有抗性的细胞系。有趣的是，这两种细胞系仍然表现出依赖于Fv Env的荧光逆转录病毒颗粒的附着，这意味着潜在的结合后阻断可能是由于缺乏假定的Fv进入辅助因子。这些细胞系可能最终导致目前未知的无处不在的细胞进入受体(S)的识别。
The foamy virus (FV) replication cycle displays several unique features, which set them apart from orthoretroviruses. First, like other B/D type orthoretroviruses, FV capsids preassemble at the centrosome, but more similar to hepadnaviruses, FV budding is strictly dependent on cognate viral glycoprotein coexpression. Second, the unusually broad host range of FV is thought to be due to use of a very common entry receptor present on host cell plasma membranes, because all cell lines tested in vitro so far are permissive. In order to take advantage of modern fluorescent microscopy techniques to study FV replication, we have created FV Gag proteins bearing a variety of protein tags and evaluated these for their ability to support various steps of FV replication. Addition of even small N-terminal HA-tags to FV Gag severely impaired FV particle release. For example, release was completely abrogated by an N-terminal autofluorescent protein (AFP) fusion, despite apparently normal intracellular capsid assembly. In contrast, C-terminal Gag-tags had only minor effects on particle assembly, egress and particle morphogenesis. The infectivity of C-terminal capsid-tagged FV vector particles was reduced up to 100-fold in comparison to wild type; however, infectivity was rescued by coexpression of wild type Gag and assembly of mixed particles. Specific dose-dependent binding of fluorescent FV particles to target cells was demonstrated in an Env-dependent manner, but not binding to target cell-extracted- or synthetic- lipids. Screening of target cells of various origins resulted in the identification of two cell lines, a human erythroid precursor- and a zebrafish- cell line, resistant to FV Env-mediated FV- and HIV-vector transduction. We have established functional, autofluorescent foamy viral particles as a valuable new tool to study FV - host cell interactions using modern fluorescent imaging techniques. Furthermore, we succeeded for the first time in identifying two cell lines resistant to Prototype Foamy Virus Env-mediated gene transfer. Interestingly, both cell lines still displayed FV Env-dependent attachment of fluorescent retroviral particles, implying a post-binding block potentially due to lack of putative FV entry cofactors. These cell lines might ultimately lead to the identification of the currently unknown ubiquitous cellular entry receptor(s) of FVs.
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影响因子：
5.4
作者：
BANFIELD, BW;LEDUC, Y;TUFARO, F
通讯作者：
TUFARO, F
影响因子：
5.3
作者：
DUBRIDGE, RB;TANG, P;CALOS, MP
通讯作者：
CALOS, MP
影响因子：
5.4
作者：
Lindemann, D;Rethwilm, A
通讯作者：
Rethwilm, A
DOI：
10.1016/0302-4598(88)80013-8
发表时间：
1988-06-01
期刊：
BIOELECTROCHEMISTRY AND BIOENERGETICS
影响因子：
--
作者：
DIMITROV, DS;ANGELOVA, MI
通讯作者：
ANGELOVA, MI
DOI：
10.1073/pnas.0504812102
发表时间：
2005-10-25
影响因子：
11.1
作者：
Larson, DR;Johnson, MC;Volker, VM
通讯作者：
Volker, VM