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Biophysical Screens Identify Fragments That Bind to the Viral DNA-Binding Proteins EBNA1 and LANA
Troy E Messick1, Lois Tolvinski1, Edward R Zartler2
1The Wistar Institute, 3601 Spruce Street, Philadelphia, PA 19104, USA.
Abstract:
The human gamma-herpesviruses Epstein-Barr virus (EBV) (HHV-4) and Kaposi's sarcoma-associated herpesvirus (KSHV) (HHV-8) are responsible for a number of diseases, including various types of cancer. Epstein-Barr nuclear antigen 1 (EBNA1) from EBV and latency-associated nuclear antigen (LANA) from KSHV are viral-encoded DNA-binding proteins that are essential for the replication and maintenance of their respective viral genomes during latent, oncogenic infection. As such, EBNA1 and LANA are attractive targets for the development of small-molecule inhibitors. To this end, we performed a biophysical screen of EBNA1 and LANA using a fragment library by saturation transfer difference (STD)-NMR spectroscopy and surface plasmon resonance (SPR). We identified and validated a number of unique fragment hits that bind to EBNA1 or LANA. We also determined the high-resolution crystal structure of one fragment bound to EBNA1. Results from this screening cascade provide new chemical starting points for the further development of potent inhibitors for this class of viral proteins.
Insights
Researchers screened for small-molecule inhibitors targeting Epstein-Barr nuclear antigen 1 (EBNA1) and latency-associated nuclear antigen (LANA) from human gamma-herpesviruses. This study identified novel fragment hits, offering new chemical starting points for developing antiviral therapies against EBV and KSHV infections.
Area of Science:
- Virology
- Molecular Biology
- Drug Discovery
Background:
- Human gamma-herpesviruses Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV) are linked to various cancers.
- Viral proteins EBNA1 (EBV) and LANA (KSHV) are crucial for viral genome maintenance during latent infection and are potential therapeutic targets.
Purpose of the Study:
- To identify small-molecule inhibitors targeting EBNA1 and LANA.
- To discover novel chemical starting points for developing antiviral drugs against EBV and KSHV.
Main Methods:
- Biophysical screening of a fragment library using saturation transfer difference (STD)-NMR spectroscopy.
- Surface plasmon resonance (SPR) assays for fragment validation.
- High-resolution crystal structure determination of a fragment bound to EBNA1.
Main Results:
- Identified and validated unique fragment hits that bind to EBNA1 or LANA.
- Determined the crystal structure of an EBNA1-fragment complex.
- Provided new chemical scaffolds for inhibitor development.
Conclusions:
- The identified fragments represent promising starting points for developing small-molecule inhibitors against EBNA1 and LANA.
- This work facilitates the development of novel therapeutics targeting oncogenic human gamma-herpesviruses.
