Polypyrimidine tract-binding protein interacts with the 3' stem-loop region of Japanese encephalitis virus negative-strand RNA.
The 3' stem-loop (SL) region of positive- and negative-strand RNA of Japanese encephalitis virus (JEV), like that of other flaviviruses, may function as cis-acting signals during RNA replication. In order to demonstrate the specific interaction between JEV 3' SL regions and BHK-21 cellular proteins, we performed gel mobility shift assay and UV-induced cross-linking assay. We identified seven cellular proteins of 110, 87, 67, 45, 38, 34, and 30 kDa that bound to the (+)3' SL RNA, and eight cellular proteins of 138, 110, 87, 67, 55, 52, 38, and 34 kDa that bound to the (-)3' SL RNA. The 55 kDa protein was identified as the polypyrimidine tract-binding (PTB) protein by immunoprecipitation assay. These data suggest that the 3' SL regions of JEV-RNA of both polarities may act as recruiting signals for the components of viral replication complexes including host cell-derived PTB protein.
Kim
Seong Man
SM
Department of Biology, College of Sciences, Kyung Hee University, Dongdaemun-gu, Seoul 130-701, South Korea.
Jeong
Yong Seok
YS
D020413:3' Untranslated Regions / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D024202:Electrophoretic Mobility Shift Assay; D004664:Encephalitis Virus, Japanese / Q000502:physiology*; D047468:Immunoprecipitation; D008970:Molecular Weight; D009690:Nucleic Acid Conformation; D038941:Polypyrimidine Tract-Binding Protein / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism*
D020413:3' Untranslated Regions; D011506:Proteins; D012367:RNA, Viral; D038941:Polypyrimidine Tract-Binding Protein