Crystal Structure of the Gamma-2 Herpesvirus LANA DNA Binding Domain Identifies Charged Surface Residues Which Impact Viral Latency

PLoS Pathogens

Chantal Beauchemin¹  Shijun Li¹  Kenneth M. Kaye¹  Thomas R. Schneider²  Sofia A. Cerqueira³  Marta Pires de Miranda³  J. Pedro Simas³  Lénia Rodrigues³  Maria A. Carrondo⁴  Colin E. McVey⁴  Bruno Correia⁴  Rajesh Ponnusamy⁴ 

[1] Departments of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, United States of America;EMBL c/o DESY, Hamburg, Germany;Instituto de Microbiologia e Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisboa, Portugal;Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal

关键词: Viral persistence;    latency;    DNA-binding proteins;    Glutathione chromatography;    B cells;    DNA structure;    Dimers (Chemical physics);    Crystal structure;    Phosphates;   

DOI  :  10.1371/journal.ppat.1003673

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Latency-associated nuclear antigen (LANA) mediates γ2-herpesvirus genome persistence and regulates transcription. We describe the crystal structure of the murine gammaherpesvirus-68 LANA C-terminal domain at 2.2 Å resolution. The structure reveals an alpha-beta fold that assembles as a dimer, reminiscent of Epstein-Barr virus EBNA1. A predicted DNA binding surface is present and opposite this interface is a positive electrostatic patch. Targeted DNA recognition substitutions eliminated DNA binding, while certain charged patch mutations reduced bromodomain protein, BRD4, binding. Virus containing LANA abolished for DNA binding was incapable of viable latent infection in mice. Virus with mutations at the charged patch periphery exhibited substantial deficiency in expansion of latent infection, while central region substitutions had little effect. This deficiency was independent of BRD4. These results elucidate the LANA DNA binding domain structure and reveal a unique charged region that exerts a critical role in viral latent infection, likely acting through a host cell protein(s).

【 授权许可】

CC BY