NSF Org:	CHE Division Of Chemistry
Recipient:	RECTOR & VISITORS OF THE UNIVERSITY OF VIRGINIA
Initial Amendment Date:	June 2, 2020
Latest Amendment Date:	June 2, 2020
Award Number:	2029895
Award Instrument:	Standard Grant
Program Manager:	Catalina Achim cachim@nsf.gov  (703)292-2048 CHE  Division Of Chemistry MPS  Directorate for Mathematical and Physical Sciences
Start Date:	June 15, 2020
End Date:	May 31, 2022 (Estimated)
Total Intended Award Amount:	$150,000.00
Total Awarded Amount to Date:	$150,000.00
Funds Obligated to Date:	FY 2020 = $150,000.00
History of Investigator:	Bryan Berger (Principal Investigator) bwb2k@virginia.edu
Recipient Sponsored Research Office:	University of Virginia Main Campus 1001 EMMET ST N CHARLOTTESVILLE VA  US  22903-4833 (434)924-4270
Sponsor Congressional District:	05
Primary Place of Performance:	University of Virginia Main Campus 102 Engineers' Way Charlottesville VA  US  22904-4741
Primary Place of Performance Congressional District:	05
Unique Entity Identifier (UEI):	JJG6HU8PA4S5
Parent UEI:
NSF Program(s):	COVID-19 Research
Primary Program Source:	010N2021DB R&RA CARES Act DEFC N
Program Reference Code(s):	096Z, 7916
Program Element Code(s):	158Y00
Award Agency Code:	4900
Fund Agency Code:	4900
Assistance Listing Number(s):	47.049

Note: This Award includes Coronavirus Aid, Relief, and Economic Security (CARES) Act funding.

ABSTRACT

With this award, the Chemistry of Life Processes Program in the Chemistry Division and the Chemical and Biochemical Engineering Program in the Chemical, Bioengineering, Environmental and Transport Systems Division are funding Dr. Bryan Berger (University of Virginia) and Dr. Jeffery Klauda (University of Maryland) to investigate two proteins named ORF7a and OR7b from the COVID19 virus that have been implicated in how harmful the virus is to its host, e.g. the human cells. The research will focus on how these two proteins form larger protein complexes that in turn affect the interactions between the virus and the infected cells and influence the immune response of the host. The research informs the development of peptides that could be used to probe the viral propagation. The research is based on the use of a combination of computational and experimental methods. Dr. Berger and Dr. Klauda distribute to the scientific community free of charge through Addgene the plasmids and associated protocols developed for this project, thus enabling the global scientific community that works on finding a solution to the current pandemic and to minimizing the possibility of future outbreaks to quickly use the outcomes of their research. This work will provide training for post-doctoral fellows working on critical challenges using state-of-the-art experimental and computational methods. The results of the research will be disseminated by the team to the greater community through conferences and workshops at University of Virginia and University of Maryland and through publications. The researchers also plan to inform and educate students on possible mechanisms of virus transmission and prevention by participation in existing outreach programs at their Institutions.

This research project seeks to understand the basis of specificity for transmembrane and juxtamembrane oligomerization of ORF7a with BST-2 and for homooligomerization of ORF7b. Using bacterial transcriptional assays for membrane protein dimerization based on the E. coli AraC protein (AraTM and DN-AraTM assays), the researches determine specific amino acid residues and structural motifs responsible for the protein oligomerization in bacterial membranes. This knowledge informs computational models for formation of BST-2/ORF7a heterooligomers and ORF7b homooligomers. In turn, the computational models are used to make critical new predictions of sequences for transmembrane peptides that could influence protein-protein interactions involving ORF7a and ORF7b. These predictions and the properties of the peptides are tested by synthesizing peptide libraries and using AraTM, DN-AraTM, and mammalian, cell-based fluorescence resonance energy transfer assays. Validation of candidate sequences are achieved using mammalian cell-based assays for BST-2 function and apoptosis. The results of these studies could provide high-resolution, experimentally validated models for OR7a and ORF7b homo and heterooligomerization, as well as peptide sequences that can be used to probe the roles of ORF7a and ORF7b in viral propagation in vivo.

This grant is being awarded using funds made available by the Coronavirus Aid, Relief, and Economic Security (CARES) Act supplement allocated to MPS and ENG.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

PROJECT OUTCOMES REPORT

Disclaimer

This Project Outcomes Report for the General Public is displayed verbatim as submitted by the Principal Investigator (PI) for this award. Any opinions, findings, and conclusions or recommendations expressed in this Report are those of the PI and do not necessarily reflect the views of the National Science Foundation; NSF has not approved or endorsed its content.

This NSF grant focused on researching two accessory proteins produced by the SARS-CoV-2 virus that has been suggested to play an important role in the level of infection. We were able to probe how one accessory protein (ORF7a) interacts with a native protein in humans (BST-2) to suppress its antiviral activity. Our combined experimental and computational approach clearly show the importance of how the ORFa/BST-2 dimer is stabilized and how a natural mutation in the human population results in more preference of BST-2 to bind to ORF7a potentially resulting in a reduced cellular antivirial activity. We have also made some initial headway into investigating potential peptide inhibitors for the ORF7a/BST-2 interface as a potential drug target. Overall, this project produced a submitted peer-reviewed research paper with another to be submitted early 2022 and trained 2 postdoctoral fellows. Specific areas of research and findings are as follows:

• Molecular dynamics (MD) simulations and experiments probed important ORF7a/BST-2 contacts that support the stability of this dimer that suppresses a cell’s natural antivirial response 
• MD and experiments probed the homodimer interface of ORF7b (another accessory protein made by SARS-CoV-2)
• Developed a library of potential peptide inhibitors for the ORF7a/BST-2 interface
• Research results have been disseminated to the scientific community in submitted peer-reviewed publications and talks at national conferences and national/international universities.
• Trained two postdoctoral researchers
• This research was also presented to undergraduates during classes to provide context of class material to the pandemic
• The research was also presented to the public in the form of an invited podcast by NBA Hall of Fame basketball player Ralph Sampson

 

Last Modified: 12/23/2022
Modified by: Bryan W Berger

Please report errors in award information by writing to: awardsearch@nsf.gov.