Folding of the SARS‐CoV‐2 RNA Polymerase by the Cytosolic Chaperonin CCT

The ongoing Covid‐19 pandemic, caused by the highly transmissible SARS‐CoV‐2 virus, constitutes the worst global public health crisis of the past century. Although effective vaccines have been developed, administration has been slow, new variants continue to emerge, and there remains a lack of effective antivirals to treat severe cases. Thus, there remains a significant need to understand the mechanisms of SARS‐CoV‐2 infection and replication to identify new potential therapeutic targets. To this end, we investigated whether SARS‐CoV‐2 might depend on the host cell chaperone system, particularly the essential cytosolic chaperonin, CCT, to fold or assemble any of its proteins. CCT has previously been shown to be required for the replication of several other viruses including reoviruses and zika virus. We screened likely candidates among the SARS‐CoV‐2 proteins for co‐immunoprecipitation with CCT and identified an interaction with the RNA‐dependent RNA polymerase, Nsp12. To confirm Nsp12 was a substrate, we depleted cells of CCT, then transfected with Nsp12 and observed a 40% decrease in Nsp12 expression in CCT‐depleted cells compared to the control. This decrease is consistent with what we have observed for other known CCT substrates. Additionally, CCT‐depleted cells infected with live SARS‐CoV‐2 produced a 50% decrease in viral titer compared to controls, indicating that CCT is important for viral replication. A preliminary cryo‐EM structure of the Nsp12‐CCT complex shows a very large mass identifiable as Nsp12 inside of CCT that extends from the equatorial domains up and out through one of the two folding cavities. The structure stands to both reveal an important step in SARS‐CoV‐2 replication and to answer a long‐standing question regarding how CCT can accommodate very large substrates.

Electronic case reporting (eCR) of COVID-19 to public health: implementation perspectives from the Minnesota Department of Health.

Electronic case reporting (eCR) is the automated generation and transmission of case reports from electronic health records to public health for review and action. These reports (electronic initial case reports: eICRs) adhere to recommended exchange and terminology standards. eCR is a partnership of the Centers for Disease Control and Prevention (CDC), Association of Public Health Laboratories (APHL) and Council of State and Territorial Epidemiologists (CSTE). The Minnesota Department of Health (MDH) received eICRs for COVID-19 from April 2020 (3 sites, manual process), automated eCR implementation in August 2020 (7 sites), and on-boarded ∼1780 clinical units in 460 sites across 6 integrated healthcare systems (through March 2022). Approximately 20 000 eICRs/month were reported to MDH during high-volume timeframes. With increasing provider/health system implementation, the proportion of COVID-19 cases with an eICR increased to 30% (March 2022). Evaluation of data quality for select demographic variables (gender, race, ethnicity, email, phone, language) across the 6 reporting health systems revealed a high proportion of completeness (>80%) for half of variables and less complete data for rest (ethnicity, email, language) along with low ethnicity data (<50%) for one health system. Presently eCR implementation at MDH includes only one EHR vendor. Next steps will focus on onboarding other EHRs, additional eICR data extraction/utilization, detailed analysis, outreach to address data quality issues, and expanding to other reportable conditions.