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Specific interaction between coronavirus leader RNA and nucleocapsid protein
S A Stohlman1, R S Baric, G N Nelson
1Department of Neurology, University of Southern California School of Medicine, Los Angeles 90033.
Journal of Virology
|November 1, 1988
Summary
The mouse hepatitis virus nucleocapsid protein (N) binds specifically to conserved sequences in the viral leader RNA. This interaction is crucial for the unique discontinuous RNA transcription process in coronaviruses.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Coronaviruses, including mouse hepatitis virus (MHV), possess a unique RNA transcriptional process.
- The nucleocapsid protein (N) of MHV is known to interact with viral RNAs.
Purpose of the Study:
- To investigate the specific interaction between the MHV nucleocapsid protein (N) and virus-specific RNAs.
- To identify the RNA sequences involved in this high-affinity binding.
Main Methods:
- Northwestern blotting using radiolabeled viral RNAs and host cell competitor RNA.
- In vitro transcription to prepare RNA probes for binding site analysis.
Main Results:
- Demonstrated a high-affinity interaction between the MHV N protein and virus-specific RNAs.
- Identified specific binding of the N protein to sequences within the conserved leader RNA.
- Pinpointed the binding site to nucleotides 56-65 at the 3' end of the leader RNA.
Conclusions:
- The MHV N protein specifically binds to a conserved region of the leader RNA.
- This interaction likely plays a significant role in the discontinuous, nonprocessive RNA transcription characteristic of coronaviruses.