A new study maps SARS-CoV-2 and hosts protein interactions

A new study was released as an introduction to the bioRxiv* the server has developed a systematic map detailing the mechanical interactions and interactions between severe acute respiratory coronavirus 2 (SARS-CoV-2) syndrome and its host cell.

The research finds host proteins that interact with SARS-CoV-2 proteins to enhance functions such as immune signaling, inflammation, protein normalization, and membrane trade-off.

The findings could help to understand how SARS-CoV-2 interacts with human proteins and create potential entry ports for therapeutic targets to break this relationship. .

How they did it

The team used two independent Y2H variants with His3 selection with growth-based readings and a green fluorescent protein system in which the GFP reporter gene is measured in fluorescence-activated cell resolution.

Cloning of SARS-CoV-2 proteins was created for Y2H_HIS3, and a codon-coded SARS-CoV-2 (ORF) open reading frame was modified for Y2H_GFP. It was then transferred into copy predatory plasmids as N-terminal fusions. To evaluate human ORFs, they collected duplicate predatory plasmids with C-terminal fusions, which covered approximately 14,000 human ORFs with two unique barcoded plasmids.

Using Y2H_HIS3, the researchers screened for SARS-CoV-2 ORF against 17,412 ORFs, with all proteins screened and matched as both feed and predatory beverages. They ordered plasmids to confirm that the viral and host proteins interacted with each other. Paired interactions that received a positive score at least twice and did not show growth in selected media without the predatory plasmid were identified as bona fide Y2H Interactions.

In total, 119 viral-host interactions used 14 viral proteins and 93 human host proteins. The researchers used these results to create an interactive binary SARS-CoV-2 human map called HuSCI_HIS3.

The second strategy uses the Y2H_GFP screening evaluated 14,627 human ORFs with two unrelated barcoded plasmids attached to 27,671 barcoded-like rays. Each strain was scraped and attached to 28 viral OFGs for a possible interaction with each other for 409,556 predatory feed combinations that were able to be represented by 2,269,022 unique barcoded diploid rays. The screening found 27 intraviral interactions using 19 viral proteins.

Together, the research team identified 205 direct viral interactions and 27 intraviral binary interactions among 171 cell-host proteins and 19 SARS-CoV-2 proteins.

Further tests showed that their network of 205 host viruses and 27 intraviral interactions was of sufficiently high quality. The authors note a series of observations describing a high-quality network, including the enrichment of infection-related host-viral interactions, 62 of interactions critical of the targets for other viruses, a large number of physically targeted proteins altered their phosphorylation status on infection, and translocation of intraviral interactions between the two networks.

The researchers consider SARS-CoV-2 spreading across species due to increased less reliable and weaker virus-host protein interactions based on their network results.

With the viral-host interactions, the team re-examined which cell processes from host proteins developed. They found an enrichment of viral trade, including central vesicle transport to the plasma membrane and Golgi network and immune regulation.

Therefore, in addition to its actions in viral reproduction, interaction analysis suggests a role for NSP16 in virion production and release, ”the team wrote.

Although there were differences in activity consistency, the two networks showed complementary but consistent levels of action on the interaction. The researchers suggest that the findings could provide more insight into how the virus controls immune and ubiquitination processes.

They also found that TRIM protein family members, namely TRIM2, 3, 27, 32, 50, and 54, were interaction targets for viral NSP16 and NSP14.

Taken together, the identified interactions reveal direct pathways by which SARS-CoV-2 may target both type I IFN pathways (and thus tissue-protective protective signals). antiviral) and inflammatory cytokine signaling. ”

Considering the functional disruption of cytotoxic lymphocytes observed during immune responses against SARS-CoV-2, the researchers examined the mechanism behind the action. They found that NSP6 interacted with CD40 and CD2,7, which regulate cytotoxic lymphocytes. The researchers suggest that NSP6 may act as a potential regulator in the development of T cell and other COVID-19 signals.

SARS-CoV-2 viral proteins, specifically ORF3D, ORF5, and ORF9C, also targeted the UBQLN1 / 2 host protein associated with ubiquitin-mediated contamination.

Based on the findings, the researchers expect that their map of SARS-CoV-2 protein interactions with host proteins will provide a greater understanding of viral multiplication and create opportunities for therapeutic interventions for pandemics in the future. future.

* Important message

bioRxiv publish preliminary scientific reports that are not peer-reviewed and, therefore, should not be seen as final, guiding health-related clinical practice / behavior, or be treated as information established.