A Human Cytomegalovirus-Encoded microRNA Regulates Expression of Multiple Viral Genes Involved in Replication

PLoS Pathogens

Deborah H Spector¹  Elizabeth A White²  Jay Nelson³  Heather Meyers³  Finn Grey³ 

[1] Cellular and Molecular Medicine, University of California San Diego, La Jolla, California, United States of America;Department of Pathology, Harvard Medical School, Boston, Massachusetts, United States of America;Vaccine and Gene Therapy Institute, Oregon Health Sciences University, Portland, Oregon, United States of America

关键词: MicroRNAs;    Viral replication;    Gene expression;    Human cytomegalovirus;    Gene regulation;    Viral gene expression;    DNA replication;    Transfection;   

DOI  :  10.1371/journal.ppat.0030163

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Although multiple studies have documented the expression of over 70 novel virus-encoded microRNAs (miRNAs), the targets and functions of most of these regulatory RNA species are unknown. In this study a comparative bioinformatics approach was employed to identify potential human cytomegalovirus (HCMV) mRNA targets of the virus-encoded miRNA miR-UL112-1. Bioinformatics analysis of the known HCMV mRNA 3′ untranslated regions (UTRs) revealed 14 potential viral transcripts that were predicted to contain functional target sites for miR-UL112-1. The potential target sites were screened using luciferase reporters that contain the HCMV 3′UTRs in co-transfection assays with miR-UL112-1. Three of the 14 HCMV miRNA targets were validated, including the major immediate early gene encoding IE72 (UL123, IE1), UL112/113, and UL120/121. Further analysis of IE72 regulation by miR-UL112-1 with clones encoding the complete major immediate early region revealed that the IE72 3′UTR target site is necessary and sufficient to direct miR-UL112-1-specific inhibition of expression in transfected cells. In addition, miR-UL112-1 regulation is mediated through translational inhibition rather than RNA degradation. Premature expression of miR-UL112-1 during HCMV infection resulted in a significant decrease in genomic viral DNA levels, suggesting a functional role for miR-UL112-1 in regulating the expression of genes involved in viral replication. This study demonstrates the ability of a viral miRNA to regulate multiple viral genes.

【 授权许可】

CC BY