Compartmentalization-aided interaction screening reveals extensive high-order complexes within the SARS-CoV-2...

Area of Science:

• Virology
• Molecular Biology
• Biochemistry

Background:

• Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) relies on complex replication/transcription complexes (RTCs) for infection.
• Understanding the interactions within these viral protein networks is crucial for developing effective treatments.

Purpose of the Study:

• To develop and apply an innovative cell-based screening strategy, compartmentalization of protein-protein interactions in cells (CoPIC), to map the interaction network of SARS-CoV-2 RTC-related proteins.
• To identify binary and ternary protein complexes involved in viral replication, transcription, and translation.

Main Methods:

• Development of a novel cell-based interaction screening strategy named compartmentalization of protein-protein interactions in cells (CoPIC).
• Application of CoPIC to screen and map interactions among 14 key viral proteins involved in SARS-CoV-2 replication and transcription.

Main Results:

• Identification of 47 binary protein-protein interactions among 14 RTC-related viral proteins.
• Discovery of extensive ternary complexes, suggesting the existence of higher-order protein structures within the RTC.
• Demonstration of CoPIC as an efficient and robust method for screening protein interactions in situ.

Conclusions:

• The study presents an effective CoPIC strategy for dissecting viral protein interaction networks.
• A complex interaction network among SARS-CoV-2 RTC factors has been elucidated.
• These findings offer new avenues for understanding SARS-CoV-2 biology and designing therapeutic interventions against COVID-19.