Role of CD151, A tetraspanin, in porcine reproductive and respiratory syndrome virus infection.
BACKGROUND
Porcine reproductive and respiratory syndrome virus (PRRSV) is a RNA virus causing respiratory and reproductive diseases in swine. The susceptibility for PRRSV varies between the different breeds of swine. In cell culture, PRRSV virus can be propagated in primary porcine alveolar macrophages and some African green monkey kidney cell lines, such as MARC-145 cells. Previous studies have shown that 3' untranslated region (UTR) RNAs of the arteriviruses play an important role in the replication of the virus through interactions with cellular proteins. To better understand the differences in the replication capability of PRRSV in different cell lines, we sought to identify the host cellular proteins interacting with PRRSV 3' UTR RNA. We constructed a cDNA library of MARC-145 cell line in lambda ZAP Express vector and screened the library with the positive sense 3' UTR RNA of PRRSV.


RESULTS
We found that CD151, a host cellular protein, interacting with PRRSV 3' UTR RNA. The specificity of the interaction between CD151 and PRRSV 3' UTR RNA was examined by gel shift assay as well as North-Western hybridization. The transfection of CD151 expression clone into BHK-21 rendered these cells susceptible to PRRSV infection, and the transfection of siRNA against CD151 into MARC-145 significantly reduced the level of PRRSV infection. Also, anti-CD151 antibody treatment to MARC-145 completely blocked PRRSV infection.


CONCLUSIONS
Based on our results, we suggest that CD151 should cooperate in PRRSV infection in vitro in MARC-145 and BHK-21 cells.
Shanmukhappa
Kumar
K
Division of Pediatric Gastroenterology, Hepatology and Nutrition, Cincinnati Children's Hospital Medical Center, University of Cincinnati. Cincinnati, OH 42229, USA. shatm2@cchmc.org
Kim
Jeong-Ki
JK
Kapil
Sanjay
S
D020413:3' Untranslated Regions / Q000378:metabolism*; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D002417:Cattle; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D024202:Electrophoretic Mobility Shift Assay; D008647:Mesocricetus; D019316:Porcine respiratory and reproductive syndrome virus / Q000254:growth & development*; D011485:Protein Binding; D034741:RNA, Small Interfering; D012367:RNA, Viral / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism; D013552:Swine; D060185:Tetraspanin 24; D014162:Transfection
D020413:3' Untranslated Regions; D015703:Antigens, CD; D034741:RNA, Small Interfering; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D060185:Tetraspanin 24