Analysis of SARS-CoV receptor activity of ACE2 orthologs.
Analysis of SARS-CoV receptor activity of ACE2 orthologs.
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ACE2 直向同源物的 SARS-CoV 受体活性分析。
DOI：
10.1007/978-0-387-33012-9_46
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发表时间：
2006
影响因子：
--
通讯作者：
Wentworth,DavidE
中科院分区：
文献类型：
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作者：
Olivieri,EmilyR;Heller,LindseyK;Gillim-Ross,Laura;Wentworth,DavidE
1传播的SARS-CoV可能是负责爆发，病毒感染多个物种。人血管紧张素转换酶2（ACE 2）是SARS冠状病毒的功能性受体。2冠状病毒刺突蛋白-受体相互作用是种属特异性的主要决定因素，病毒基因组RNA的转染或受体在非允许细胞系中的表达通常会导致生产性感染。我们的假设是ACE 2的物种特异性差异在SARS-CoV感染中是重要的，并且ACE 2直系同源物的分析将允许识别对病毒进入至关重要的受体区域。我们分析了来自许多物种的细胞系对SARS-CoV的易感性。来自人、猴和貂的细胞系对SARS-CoV是允许的，并且在所有这些细胞系中检测到ACE 2 RNA转录物。ACE 2 RNA也在非允许的狗（Canis familiaris）和鸡（Gallus gallus）细胞中检出。我们使用人ACE 2（hACE 2）和小鼠ACE 2之间的保守区域，通过RT-PCR和3 'RACE扩增来自不同物种的ACE 2的区域。序列分析表明，犬ACE 2与hACE 2的核苷酸同源性为87%，氨基酸同源性为81%，鸡ACE 2与hACE 2的核苷酸同源性为79%，氨基酸同源性为62%。从犬和鸡的cDNA克隆中获得ACE 2开放阅读框，并将其亚克隆到真核表达系统中，以分析其作为SARS-CoV受体的功能。
1 transmission of SARS-CoV was likely responsible for the outbreak, and the virus infects multiple species. Human angiotensin-converting enzyme 2 (ACE2) is a functional receptor for SARS-CoV. 2 Coronavirus spike-receptor interactions are major determinants of species specificity, and transfection of viral genomic RNA or expression of receptors in nonpermissive cell lines usually results in productive infection. Our hypothesis is that species-specific differences in ACE2 are important in SARS-CoV infection, and analysis of ACE2 orthologs will permit the identification of regions of the receptor that are critical for virus entry. We analyzed cell lines that were derived from numerous species for their susceptibility to SARS-CoV. Cell lines derived from human, monkey, and mink were permissive to SARS-CoV and ACE2 RNA transcripts were detected in all of these cell lines. ACE2 RNA was also detected in nonpermissive dog (Canis familiaris) and chicken (Gallus gallus) cells. We used regions conserved between human ACE2 (hACE2) and mouse ACE2 to amplify regions of ACE2 from diverse species by RT-PCR and 3’RACE. Sequence analysis demonstrated that dog ACE2 had 87% nucleotide identity and 81% amino acid identity with hACE2, and chicken ACE2 has 79% nucleotide identity and 62% amino acid identity with hACE2. The ACE2 open reading frames from dog and chicken cDNA clones were subcloned into eukaryotic expression to analyze their function as SARS-CoV receptors.