a human cytomegalovirus-encoded microrna regulates expression of multiple viral genes involved in replication人类cytomegalovirus-encoded微rna调节表达多个病毒基因的复制.pdfVIP

a human cytomegalovirus-encoded microrna regulates expression of multiple viral genes involved in replication人类cytomegalovirus-encoded微rna调节表达多个病毒基因的复制.pdf

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a human cytomegalovirus-encoded microrna regulates expression of multiple viral genes involved in replication人类cytomegalovirus-encoded微rna调节表达多个病毒基因的复制

A Human Cytomegalovirus-Encoded microRNA Regulates Expression of Multiple Viral Genes Involved in Replication 1* 1 2 3 1 Finn Grey , Heather Meyers , Elizabeth A. White , Deborah H. Spector , Jay Nelson 1 Vaccine and Gene Therapy Institute, Oregon Health Sciences University, Portland, Oregon, United States of America, 2 Department of Pathology, Harvard Medical School, Boston, Massachusetts, United States of America, 3 Cellular and Molecular Medicine, University of California San Diego, La Jolla, California, United States of America 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 39 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 39UTRs 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 39UTR 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 gen

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