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1、Heparan Sulfate Is a Selective Attachment Factor for the Avian Coronavirus Infectious Bronchitis Virus Beaudette (El heparn sulfato es un factor de unin para la cepa Beaudette del virus de bronquitis infecciosa) Author(s): Ikenna G. Madu, Victor C. Chu, Hwajin Lee, Andrew D. Regan, Beverley E. Bauma
2、n and Gary R. Whittaker Source: Avian Diseases, Vol. 51, No. 1 (Mar., 2007), pp. 45-51 Published by: American Association of Avian Pathologists Stable URL: http:/www.jstor.org/stable/4493167 . Accessed: 20/06/2014 21:08 Your use of the JSTOR archive indicates your acceptance of the Terms Accepted 14
3、 September 2006 SUMMARY. The avian coronavirus infectious bronchitis virus (IBV) strain Beaudette is an embryo-adapted virus that has extended species tropism in cell culture. In order to understand the acquired tropism of the Beaudette strain, we compared the S protein sequences of several IBV stra
4、ins. The Beaudette strain was found to contain a putative heparan sulfate (HS)-binding site, indicating that the Beaudette virus may use HS as a selective receptor. To ascertain the requirements of cell-surface HS for Beaudette infectivity, we assayed for infectivity in the presence of soluble hepar
5、in as a competitor and determined infectivity in mutant cell lines with no HS or glycosaminoglycan expression. Our results indicate that HS plays a role as an attachment factor for IBV, working in concert with other factors like sialic acid to mediate virus binding to cells, and may explain in part
6、the extended tropism of IBV Beaudette. RESUMEN. El heparain sulfato es un factor de uni6n para la cepa Beaudette del virus de bronquitis infecciosa. La cepa Beaudette del virus de bronquitis infecciosa aviar (coronavirus), es un virus adaptado a embri6n que tiene tropismo por un amplio rango de espe
7、cies en cultivo celular. Con la finalidad de comprender el tropismo adquirido por la cepa Beaudette, se compararon las secuencias de la proteina S de varias cepas de bronquitis infecciosa aviar. Se demostr6 que la cepa Beaudette contiene un sitio de union putativo para el heparan sulfato, lo que ind
8、ica que el virus Beaudette debe utilizar el heparin sulfato como un receptor selectivo. Para confirmar que el heparan sulfato en la superficie celular es requerido para la infectividad del virus Beaudette, se analizo la infectividad del virus en la presencia de heparina soluble como competidor y se
9、determin6 la infectividad en lineas celulares mutantes que no expresan heparin sulfato o glicosaminoglicanos. Los resultados indican que el heparin sulfato juega un papel como un factor de uni6n para el virus de bronquitis infecciosa, trabajando en conjunto con otros factores como el icido sialico p
10、ara mediar la uni6n del virus con la celula y puede explicar en parte el amplio tropismo de la cepa Beaudette del virus de bronquitis infecciosa. Key words: infectious bronchitis virus, receptor, heparan sulfate, sialic acid Abbreviations: ACE2 = angiotensin-converting enzyme 2; ATCC = American Type
11、 Culture Collection; BHK = baby hamster kidney; CEK = chick embryo kidney; CHO = Chinese hamster ovary; CK = chick kidney; CoV = coronavirus; CS = chondroitin sulfate; EDTA = ethylenediaminetetraacetic acid; fAPN = feline aminopeptidase N; FFU = fluorescence focus unit; GAG = glycosaminoglycan; HS =
12、 heparan sulfate; IBV = infectious bronchitis virus; M41 = Massachusetts 41; NA = neuraminidase; NCBI = National Center for Biotechnology Information; PBS = phosphate-buffered saline; RT-PCR = reverse transcription-polymerase chain reaction; SARS = severe acute respiratory syndrome Infectious bronch
13、itis virus (IBV) is a coronavirus infecting domestic fowl. It was originally isolated in the 1930s and continues to cause major problems in the poultry industry (11). Coronaviruses are enveloped positive-stranded RNA viruses that replicate in the cytoplasm (23). They have a distinctive set of club-s
14、haped spikes on their envelope, which surrounds the viral core shell and nucleocap- sid. The coronavirus (CoV) S protein comprises a large, heavily glycosylated ectodomain that can be cleaved during biosynthesis into two subdomains (S1 and S2) by a furin-like enzyme in the Golgi apparatus (13). Not
15、all coronaviruses are cleaved, yet even without cleavage the basic subdomain structure of the S protein is conserved (25). S1 comprises the receptor-binding domain (24), and S2 comprises the fusion domain (5). The S protein of IBV is fully cleaved at the S1/S2 boundary (10), especially in chicken em
16、bryo systems. Coronaviruses show strong species and tissue tropism, and a major factor in this specificity is the virus receptor on host cells (19). Receptor binding is mediated by the viral S protein, and this has been studied in some detail for many coronaviruses, a principal exception being group 3 coronaviruses such as IBV. Group 1 coronaviruses (e.g., HCoV 229E, transmissible gastroenteritis virus, and feline infectious peritonitis virus) are well established
