Yeast based small molecule screen for inhibitors of SARS-CoV.
Yeast based small molecule screen for inhibitors of SARS-CoV.
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DOI：
10.1371/journal.pone.0028479
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Engel DA
中科院分区：
文献类型：
--
作者：
Frieman M;Basu D;Matthews K;Taylor J;Jones G;Pickles R;Baric R;Engel DA
严重急性呼吸道冠状病毒（SARS-CoV）于2002年出现，导致全球约8000例病例和10%的死亡率。SARS-CoV前体的动物宿主仍然存在，未来在人群中爆发的可能性很高。SARS-CoV木瓜蛋白酶样蛋白酶（PLP）是药物开发的一个有吸引力的目标，因为它是病毒复制所必需的，并且在人类冠状病毒中是保守的。建立了一种基于酵母的PLP活性测定方法，该方法依赖于PLP在S.啤酒。缓慢生长表型的诱导显示在60小时的时间过程中发生，为筛选通过抑制PLP功能恢复生长的小分子提供了基础。从2000名NIH成员的多样性集文库中鉴定出5种缓慢生长表型的化学抑制因子。其中之一，NSC 158362，在细胞培养中有效抑制SARS-CoV复制，对细胞无毒性作用，并且它特异性抑制SARS-CoV复制，但不抑制流感病毒复制。研究了NSC 158362对PLP蛋白酶、去泛素化酶和抗干扰素活性的影响，但该化合物不改变这些活性。另一种抑制剂NSC 158011在基于细胞的测定中显示出抑制PLP蛋白酶活性的能力。这些抑制剂的鉴定证明了基于PLP的酵母测定、抑制性化合物和SARS-CoV生物学之间的强功能联系。此外，NSC 158362的数据表明了一种抑制SARS-CoV复制的新机制，可能涉及PLP的未知活性，或者对酵母和哺乳动物细胞中修饰或绕过PLP功能的细胞靶点的直接作用。
Severe acute respiratory coronavirus (SARS-CoV) emerged in 2002, resulting in roughly 8000 cases worldwide and 10% mortality. The animal reservoirs for SARS-CoV precursors still exist and the likelihood of future outbreaks in the human population is high. The SARS-CoV papain-like protease (PLP) is an attractive target for pharmaceutical development because it is essential for virus replication and is conserved among human coronaviruses. A yeast-based assay was established for PLP activity that relies on the ability of PLP to induce a pronounced slow-growth phenotype when expressed in S. cerevisiae. Induction of the slow-growth phenotype was shown to take place over a 60-hour time course, providing the basis for conducting a screen for small molecules that restore growth by inhibiting the function of PLP. Five chemical suppressors of the slow-growth phenotype were identified from the 2000 member NIH Diversity Set library. One of these, NSC158362, potently inhibited SARS-CoV replication in cell culture without toxic effects on cells, and it specifically inhibited SARS-CoV replication but not influenza virus replication. The effect of NSC158362 on PLP protease, deubiquitinase and anti-interferon activities was investigated but the compound did not alter these activities. Another suppressor, NSC158011, demonstrated the ability to inhibit PLP protease activity in a cell-based assay. The identification of these inhibitors demonstrated a strong functional connection between the PLP-based yeast assay, the inhibitory compounds, and SARS-CoV biology. Furthermore the data with NSC158362 suggest a novel mechanism for inhibition of SARS-CoV replication that may involve an unknown activity of PLP, or alternatively a direct effect on a cellular target that modifies or bypasses PLP function in yeast and mammalian cells.
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DOI：
10.1016/s0140-6736(03)13077-2
发表时间：
2003-04-19
期刊：
Lancet (London, England)
影响因子：
--
作者：
Peiris JS;Lai ST;Poon LL;Guan Y;Yam LY;Lim W;Nicholls J;Yee WK;Yan WW;Cheung MT;Cheng VC;Chan KH;Tsang DN;Yung RW;Ng TK;Yuen KY;SARS study group
通讯作者：
SARS study group
影响因子：
5.4
作者：
Harcourt, BH;Jukneliene, D;Baker, SC
通讯作者：
Baker, SC
影响因子：
5.4
作者：
Sims, AC;Baric, RS;Pickles, RJ
通讯作者：
Pickles, RJ
DOI：
10.1016/s0140-6736(03)13413-7
发表时间：
2003-05-24
期刊：
Lancet (London, England)
影响因子：
--
作者：
Nicholls JM;Poon LL;Lee KC;Ng WF;Lai ST;Leung CY;Chu CM;Hui PK;Mak KL;Lim W;Yan KW;Chan KH;Tsang NC;Guan Y;Yuen KY;Peiris JS
通讯作者：
Peiris JS
影响因子：
5.4
作者：
Zhang, LQ;Peeples, ME;Pickles, RJ
通讯作者：
Pickles, RJ