Interactions between coronavirus nucleocapsid protein and viral RNAs: implications for viral transcription
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Implication
In vitro investigation demonstrates specific binding of coronavirus nucleocapsid protein to viral genomic and subgenomic RNAs, indicating a functional role in viral transcription and replication.
Key Points
- To investigate the direct physical interactions between the mouse hepatitis virus (MHV) nucleocapsid (N) protein and viral RNAs during infection.
- Conducted coimmunoprecipitation assays on MHV-infected cell lysates using monoclonal antibodies specific for the N protein, viral envelope glycoproteins (E1 and E2), and control sera.
- Assessed binding specificity by testing coimmunoprecipitation against host cell RNA and unrelated vesicular stomatitis virus (VSV) RNA.
- Anti-N monoclonal antibodies specifically coimmunoprecipitated MHV genomic RNA, all six subgenomic mRNAs, and negative-stranded RNA intermediates, while control antibodies and host/VSV RNAs showed no binding.
- Membrane-bound and cytosolic leader-specific RNAs longer than 65 nucleotides coimmunoprecipitated with anti-N antibodies, indicating an N-protein binding site at least 65 nucleotides from the 5' end.
Cite This Study
Baric et al. (1988) studied this question.