Cross-species conservation of episome maintenance provides a basis for in vivo investigation of Kaposi's sarcoma herpesvirus LANA

PLoS Pathogens

Edward J. Usherwood¹  Chantal Beauchemin²  Kenneth M. Kaye²  Min Tan²  Aline C. Habison²  Sofia A. Cerqueira³  Marta Pires de Miranda³  J. Pedro Simas³  Colin E. McVey⁴  Bruno Correia⁴  Rajesh Ponnusamy⁴ 

[1] Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Lebanon, New Hampshire, United States of America;Departments of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts, United States of America;Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisboa, Portugal;Instituto de Tecnologia Quimica e Bioliogica Antonio Xavier, Universidade Nova de Lisboa, Oeiras, Portugal

关键词: B cells;    Kaposi's sarcoma-associated herpesvirus;    Spleen;    Polymerase chain reaction;    Transfection;    Viral persistence;    latency;    Viral replication;    DNA replication;   

DOI  :  10.1371/journal.ppat.1006555

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Many pathogens, including Kaposi’s sarcoma herpesvirus (KSHV), lack tractable small animal models. KSHV persists as a multi-copy, nuclear episome in latently infected cells. KSHV latency-associated nuclear antigen (kLANA) binds viral terminal repeat (kTR) DNA to mediate episome persistence. Model pathogen murine gammaherpesvirus 68 (MHV68) mLANA acts analogously on mTR DNA. kLANA and mLANA differ substantially in size and kTR and mTR show little sequence conservation. Here, we find kLANA and mLANA act reciprocally to mediate episome persistence of TR DNA. Further, kLANA rescued mLANA deficient MHV68, enabling a chimeric virus to establish latent infection in vivo in germinal center B cells. The level of chimeric virus in vivo latency was moderately reduced compared to WT infection, but WT or chimeric MHV68 infected cells had similar viral genome copy numbers as assessed by immunofluorescence of LANA intranuclear dots or qPCR. Thus, despite more than 60 Ma of evolutionary divergence, mLANA and kLANA act reciprocally on TR DNA, and kLANA functionally substitutes for mLANA, allowing kLANA investigation in vivo. Analogous chimeras may allow in vivo investigation of genes of other human pathogens.

【 授权许可】

CC BY