Severe acute respiratory syndrome coronavirus nonstructural proteins 3, 4, and 6 induce double-membrane vesicles.
Severe acute respiratory syndrome coronavirus nonstructural proteins 3, 4, and 6 induce double-membrane vesicles.
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DOI：
10.1128/mbio.00524-13
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发表时间：
2013-08-13
期刊：
影响因子：
6.4
通讯作者：
Buchmeier MJ
中科院分区：
文献类型：
--
作者：
Angelini MM;Akhlaghpour M;Neuman BW;Buchmeier MJ
与其他正链RNA病毒一样，冠状病毒（CoV）可以重定向和重新排列宿主细胞膜，作为病毒基因组复制和转录机制的一部分。具体来说，冠状病毒诱导感染细胞形成双膜囊泡。尽管这些双膜囊泡已被很好地表征，但其形成背后的机制仍不清楚，包括哪些病毒蛋白负责。在这里，我们使用转染编码严重急性呼吸综合征（SARS）冠状病毒的三种跨膜非结构蛋白（nsps）全长版本的质粒构建物来检测每一种在组织培养中诱导双膜囊泡的能力。无论是全长形式还是c端截短形式，nsp3都具有膜无序性和增殖能力。Nsp3和nsp4一起工作具有使膜配对的能力。Nsp6也具有膜增殖能力，可诱导位于微管组织中心周围的核周囊泡。nsp3、nsp4和nsp6共同具有诱导双膜囊泡的能力，与在SARS冠状病毒感染的细胞中观察到的类似。这种活性似乎需要nsp3的全长形式才能发挥作用，因为在与nsp4和nsp6共表达c端截断的nsp3的细胞中没有看到双膜囊泡。虽然冠状病毒在人类中引起的大多数感染相对较轻，但2002年至2003年的SARS疫情和2012年人类冠状病毒中东呼吸综合征（MERS-CoV）的出现凸显了这些病毒造成严重病理和死亡的能力。深入了解冠状病毒如何接管宿主细胞的分子生物学，对于充分了解由这些病毒引起的任何已知和未来可能爆发的疫情至关重要。此外，由于膜重排是所有已知的正义单链RNA病毒都使用的一种策略，这项工作增加了这一知识体系，并且可能有助于开发未来的治疗方法，不仅适用于人类冠状病毒感染，也适用于其他病原体。
Coronaviruses (CoV), like other positive-stranded RNA viruses, redirect and rearrange host cell membranes for use as part of the viral genome replication and transcription machinery. Specifically, coronaviruses induce the formation of double-membrane vesicles in infected cells. Although these double-membrane vesicles have been well characterized, the mechanism behind their formation remains unclear, including which viral proteins are responsible. Here, we use transfection of plasmid constructs encoding full-length versions of the three transmembrane-containing nonstructural proteins (nsps) of the severe acute respiratory syndrome (SARS) coronavirus to examine the ability of each to induce double-membrane vesicles in tissue culture. nsp3 has membrane disordering and proliferation ability, both in its full-length form and in a C-terminal-truncated form. nsp3 and nsp4 working together have the ability to pair membranes. nsp6 has membrane proliferation ability as well, inducing perinuclear vesicles localized around the microtubule organizing center. Together, nsp3, nsp4, and nsp6 have the ability to induce double-membrane vesicles that are similar to those observed in SARS coronavirus-infected cells. This activity appears to require the full-length form of nsp3 for action, as double-membrane vesicles were not seen in cells coexpressing the C-terminal truncation nsp3 with nsp4 and nsp6. Although the majority of infections caused by coronaviruses in humans are relatively mild, the SARS outbreak of 2002 to 2003 and the emergence of the human coronavirus Middle Eastern respiratory syndrome (MERS-CoV) in 2012 highlight the ability of these viruses to cause severe pathology and fatality. Insight into the molecular biology of how coronaviruses take over the host cell is critical for a full understanding of any known and possible future outbreaks caused by these viruses. Additionally, since membrane rearrangement is a tactic used by all known positive-sense single-stranded RNA viruses, this work adds to that body of knowledge and may prove beneficial in the development of future therapies not only for human coronavirus infections but for other pathogens as well.