Identification of FBL2 as a geranylgeranylated required for hepatitis C cellular protein virus RNA replication
Identification of FBL2 as a geranylgeranylated required for hepatitis C cellular protein virus RNA replication
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DOI：
10.1016/j.molcel.2005.04.004
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发表时间：
2005-05-13
期刊：
影响因子：
16
通讯作者：
Ye, J
中科院分区：
文献类型：
--
作者：
Wang, CF;Gale, M;Ye, J
我们最近报道，丙型肝炎病毒 (HCV) RNA 复制需要一种或多种香叶基香叶基化宿主蛋白。通过结合[H-3]甲羟戊酸标记、免疫共沉淀和生物信息学搜索，我们鉴定出了 HCV RNA 复制所需的香叶基香叶基化宿主蛋白。这种蛋白质 FBL2 包含一个 F 盒结构域和一个 CAAX 基序 (CVIL)。它与 HCV 非结构蛋白 5A (NS5A) 形成稳定的免疫沉淀复合物。 FBL2 与 NS5A 的关联需要 FBL2 的 CAAX 基序，但不需要 F 盒。 F盒的缺失产生了一种显性失活蛋白，当在Huh7-K2040细胞中过度表达时，该蛋白会抑制HCV RNA的复制； NS5A 的共表达克服了这种抑制。 Huh7-HP 细胞中 siRNA 介导的 FBL2 mRNA 敲低 70%，HCV RNA 减少 65%；这种减少是通过编码 FBL2 摆动突变体的 cDNA 的表达来克服的。目前的数据表明，香叶基香叶基化的 FBL2 在对 HCV RNA 复制至关重要的反应中与 NS5A 结合。
We recently reported that Hepatitis C virus (HCV) RNA replication requires one or more geranylgeranylated host proteins. Using a combination of [H-3]mevalonate labeling, coimmunoprecipitation, and bioinformatic search, we identified a geranylgeranylated host protein required for HCV RNA replication. This protein, FBL2, contains an F box domain and a CAAX motif (CVIL). It forms a stable immunoprecipitable complex with the HCV nonstructural protein 5A (NS5A). The association of FBL2 with NS5A requires the CAAX motif of FBL2, but not the F box. Deletion of the F box created a dominant-negative protein that inhibited replication of HCV RNA when overexpressed in Huh7-K2040 cells; this inhibition was overcome by coexpression of NS5A. siRNA-mediated knockdown of FBL2 mRNA by 70% in Huh7-HP cells reduced HCV RNA by 65%; this reduction was overcome by expression of a cDNA encoding a wobble mutant of FBL2. The current data indicate that geranylgeranylated FBL2 binds to NS5A in a reaction crucial for HCV RNA replication.