Open AccessFunctional interaction between cellular p100 and the dengue virus 3' UTR
Author(s) -
Yingfeng Lei,
Yong Huang,
Zhang Hai,
Li Yu,
Mingjie Zhang,
Andrew I. Dayton
Publication year - 2010
Publication title -
journal of general virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.55
H-Index - 167
eISSN - 1465-2099
pISSN - 0022-1317
DOI - 10.1099/vir.0.028597-0
Subject(s) - biology , dengue virus , gene knockdown , untranslated region , three prime untranslated region , virology , viral replication , immunoprecipitation , rna , rna binding protein , microbiology and biotechnology , host factor , dengue fever , virus , cell culture , gene , genetics
Host factors interacting with the dengue virus (DENV) 3' UTR are involved in virus replication, but their roles remain poorly understood. We used RNA affinity capture and mass spectrometry to identify p100 as a host cellular protein associated with the DENV 3' UTR. By using RNA immunoprecipitation and confocal immunofluorescence analysis we demonstrated an interaction between p100 and the 3' UTR in DENV-infected cells. We identified the A4 region (the extensive stem-loop structure at the 3' end) as the binding site of p100 by studying deletion mutants. p100 knockdown specifically reduced the levels of viral RNA and viral protein in DENV-infected cells. Furthermore, downregulation of p100 reduced the expression of a heterologously expressed luciferase-3' UTR(DENV) mRNA in an A4-dependent manner, confirming the binding data and the effects of p100 knockdown on viral replication. These results provide evidence that p100 interacts with the 3' UTR of DENV and is required for normal DENV replication.