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Functional interaction between cellular p100 and the dengue virus 3' UTR
Yingfeng Lei1, Yong Huang, Hai Zhang
1Laboratory of Molecular Virology, Center for Biologics Evaluation and Research, Food and Drug Administration, 1401 Rockville Pike, Rockville, MD 20852, USA.
The Journal of General Virology
|December 15, 2010
Summary
Researchers identified p100 as a key host protein interacting with the dengue virus (DENV) 3' untranslated region (UTR). This interaction is crucial for DENV replication and viral RNA production.
Area of Science:
- Virology
- Molecular Biology
- Host-Pathogen Interactions
Background:
- Host factors interacting with dengue virus (DENV) 3' untranslated region (UTR) are implicated in viral replication.
- The specific roles of these host factors, however, remain largely uncharacterized.
Purpose of the Study:
- To identify and characterize host cellular proteins that interact with the DENV 3' UTR.
- To elucidate the role of identified host factors in the DENV replication cycle.
Main Methods:
- RNA affinity capture coupled with mass spectrometry to identify interacting proteins.
- RNA immunoprecipitation and confocal immunofluorescence to confirm protein-UTR interaction in infected cells.
- Deletion mutant analysis to pinpoint the specific binding site on the DENV 3' UTR.
Main Results:
- The host protein p100 was identified as a binding partner of the DENV 3' UTR.
- p100 specifically binds to the A4 region of the DENV 3' UTR.
- Knockdown of p100 significantly reduced viral RNA and protein levels in DENV-infected cells.
- Downregulation of p100 impaired the expression of a reporter mRNA containing the DENV 3' UTR in an A4-dependent manner.
Conclusions:
- p100 directly interacts with the DENV 3' UTR, specifically the A4 region.
- p100 is essential for efficient DENV replication.
- This study reveals a novel host factor critical for the DENV life cycle.
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