Structural and Biophysical Investigation of the Key Hotspots on the Surface of Epstein-Barr Nuclear Antigen 1 Essential for DNA Recognition and Pathogenesis.

Huma Farooque Hashmi,Zahid Hussain,Muhammad Waseem,Kaoshan Chen,Syed Shujait Ali

doi:10.3389/fmolb.2021.664436

Abstract

Epstein-Barr Virus (EBV) is considered the most important human pathogen due to its role in infections and cellular malignancies. It has been reported that this Oncolytic virus infects 90% world’s population. EBNA1 is required for DNA binding and survival of the virus and is considered an essential drug target. The biochemical and structural properties of this protein are known, but it is still unclear which residues impart a critical role in the recognition of dsDNA. Intending to disclose only the essential residues in recognition of dsDNA, this study used a computational pipeline to generate an alanine mutant of each interacting residue and determine the impact on the binding. Our analysis revealed that R469A, K514A, Y518A, R521A and R522A are the key hotspots for the recognition of dsDNA by the EBNA1. The dynamics properties, i.e. stability, flexibility, structural compactness, hydrogen bonding frequency, binding affinity, are altered by disrupting the protein-DNA contacts, thereby decreases the binding affinity. In particular, the two arginine substitution, R521A and R522A, significantly affected the total binding energy. Thus, we hypothesize that these residues impart a critical role in the dsDNA recognition and pathogenesis. This study would help to design structure-based drugs against the EBV infections.

Highlights

Due to the prevalent infections instigated by Herpes viruses, it is considered as an important virus in human pathogens flora
Using accession number 5T7X the structure of the Epstein-Barr Nuclear Antigen 1 (EBNA1) was retrieved
In the EBNA-DNA co-crystal structure, the three targeted amino acids are oriented toward the DNA but are too far from the nearest H-bond acceptor in the bases to form H-bonds. These results show that R469A, K514A, Y518A, R521A and R522A are required for DNA recognition and are the key hotspots for drug discovery

Summary

Introduction

Due to the prevalent infections instigated by Herpes viruses, it is considered as an important virus in human pathogens flora This diverse pathogenic flora Epstein-Barr Virus (EBV) is regarded as the most important human pathogen due to its role in infections and cellular malignancies (Bochkarev et al, 1996; Yasuda et al, 2011) It has been reported that this Oncolytic virus infects 90% world’s population. Epstein-Barr Nuclear Antigen 1 (EBNA1) acts to preserve the latent viral genome in proliferating cells (Okano, 1998). This protein is expressed in the malignant cell and sustain the proliferation (Leight and Sugden, 2000)

Methods

Results

Conclusion