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The Angiotensin Converting Enzyme 2 (ACE2) from Raccoon Dog can serve as an efficient Receptor for the Spike Protein of Severe Acute Respiratory Syndrome Coronavirus

¹ Wuhan Institute of Virology, Chinese Academy of Sciences;
² AIDS Institute, The University of Hong Kong Li Ka Shing Faculty of Medicine, Hong Kong SAR,China;
³ Department of Cell Biology and Genetics, College of Life Sciences, Peking University, Beijing,China

^* Corresponding author; email: df{at}wh.iov.cn

Raccoon dog is one of the suspected intermediated hosts of the severe acute respiratory syndrome coronavirus (SARS-CoV). In this study, the angiotensin-converting enzyme 2 (ACE2) gene of raccoon dog (rdACE2) was cloned and sequenced. Amino acid (aa) sequence of rdACE2 has an identity of 99.3%, 89.2%, 83.9%, and 80.4% to ACE2s from dog, masked palm civet, human, and bat respectively. There are six aa changes in rdACE2 compared to huACE2, and four changes compared to pcACE2, within the 18 residues of ACE2 known to make direct contact with the SARS-CoV S protein. A HeLa cell line stably express rdACE2 was established, Western blot analyses and enzyme activity assay indicated that the cell line expressed equivalent ACE2 as two previous established cell lines which express ACE2 from human and masked palm civet, respectively. HIV backboned pseudoviruses expressing spike proteins derived from human SARS-CoV or SARS-CoV-like viruses of masked palm civets and raccoon dogs were tested for their entry efficiencies on these cell lines. The results showed that rdACE2 is a more efficient receptor for human SARS-CoV, but not for the SARS-CoV-like viruses of masked palm civets and raccoon dogs, than huACE2 or pcACE2. This study provides useful data to elucidate the role of raccoon dog in SARS outbreaks.

Received 20 May 2009; accepted 19 July 2009.