Dengue virus nonstructural protein 3 redistributes fatty acid synthase to sites of viral replication and increases cellular fatty acid synthesis
Dengue virus nonstructural protein 3 redistributes fatty acid synthase to sites of viral replication and increases cellular fatty acid synthesis
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DOI：
10.1073/pnas.1010811107
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发表时间：
2010-10-05
影响因子：
11.1
通讯作者：
Randall, Glenn
中科院分区：
文献类型：
--
作者：
Heaton, Nicholas S.;Perera, Rushika;Randall, Glenn
登革病毒(DENV)通过修饰细胞膜来确定其复制位置。尽管最近已经描述了这些结构的3D结构，但人们对它们形成和扩张所需的细胞途径知之甚少。在这份报告中，我们使用集中的RNAi分析结合使用药理抑制剂的验证研究来检查DENV复制的宿主要求。这种方法确定了DENV复制所需的三条细胞途径：自噬、肌动蛋白聚合和脂肪酸生物合成。对病毒调节脂肪酸生物合成的进一步研究表明，该途径中的一个关键酶--脂肪酸合成酶(FASN)被重新定位到DENV复制的位置。DENV非结构蛋白3(NS3)负责FASN的募集，因为(I)在没有其他病毒蛋白的情况下表达的NS3与FASN共定位，以及(Ii)NS3与FASN以双杂交方式相互作用。在DENV感染的细胞中，脂肪酸的生物合成速率相应增加，从头合成的脂类优先与DENV RNA共分。最后，纯化的重组NS3在体外刺激FASN的活性。综上所述，这些实验表明DENV选择脂肪酸生物合成途径来建立其复制复合体。这项研究为DENV膜重塑提供了机制上的见解，并强调了通过靶向脂肪酸生物合成途径来抑制DENV复制的治疗药物的开发潜力。
Dengue virus (DENV) modifies cellular membranes to establish its sites of replication. Although the 3D architecture of these structures has recently been described, little is known about the cellular pathways required for their formation and expansion. In this report, we examine the host requirements for DENV replication using a focused RNAi analysis combined with validation studies using pharmacological inhibitors. This approach identified three cellular pathways required for DENV replication: autophagy, actin polymerization, and fatty acid biosynthesis. Further characterization of the viral modulation of fatty acid biosynthesis revealed that a key enzyme in this pathway, fatty acid synthase (FASN), is relocalized to sites of DENV replication. DENV nonstructural protein 3 (NS3) is responsible for FASN recruitment, inasmuch as (i) NS3 expressed in the absence of other viral proteins colocalizes with FASN and (ii) NS3 interacts with FASN in a two-hybrid assay. There is an associated increase in the rate of fatty acid biosynthesis in DENV-infected cells, and de novo synthesized lipids preferentially cofractionate with DENV RNA. Finally, purified recombinant NS3 stimulates the activity of FASN in vitro. Taken together, these experiments suggest that DENV co-opts the fatty acid biosynthetic pathway to establish its replication complexes. This study provides mechanistic insight into DENV membrane remodeling and highlights the potential for the development of therapeutics that inhibit DENV replication by targeting the fatty acid biosynthetic pathway.