Poliovirus subviral particles associated with progeny RNA in the replication complex

Area of Science:

• Virology
• Molecular Biology
• Biochemistry

Background:

• The poliovirus replication complex (RC) is crucial for viral RNA synthesis.
• Previous studies identified 5S protomer and 14S pentamer antigenic subviral particles within the RC.

Purpose of the Study:

• To investigate the interaction between subviral particles and viral RNA within the poliovirus RC.
• To determine the role of 5S/14S particles in the initial stages of poliovirus RNA encapsidation.

Main Methods:

• UV irradiation of a subcellular fraction containing the poliovirus RC.
• Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of cross-linked capsid proteins.
• Identification of cross-linked viral RNA species.

Main Results:

• 5S/14S antigenic subviral particles were successfully cross-linked to viral RNA in the RC.
• All capsid proteins (VP0, VP1, VP3) of the subviral particles were cross-linked to viral RNA.
• VP1 showed a bandshift in SDS-PAGE, and VP0 formed multiple RNA-linked multimers.
• Specifically, 36S RNA, not replicative intermediate RNA, was cross-linked to capsid proteins.

Conclusions:

• Encapsidation of poliovirus RNA initiates within the replication complex.
• The 14S pentameric subviral particle plays a key role in initiating poliovirus RNA encapsidation.