A viral microRNA down-regulates multiple cell cycle genes through mRNA 5'UTRs.
Global gene expression data combined with bioinformatic analysis provides strong evidence that mammalian miRNAs mediate repression of gene expression primarily through binding sites within the 3' untranslated region (UTR). Using RNA induced silencing complex immunoprecipitation (RISC-IP) techniques we have identified multiple cellular targets for a human cytomegalovirus (HCMV) miRNA, miR-US25-1. Strikingly, this miRNA binds target sites primarily within 5'UTRs, mediating significant reduction in gene expression. Intriguingly, many of the genes targeted by miR-US25-1 are associated with cell cycle control, including cyclin E2, BRCC3, EID1, MAPRE2, and CD147, suggesting that miR-US25-1 is targeting genes within a related pathway. Deletion of miR-US25-1 from HCMV results in over expression of cyclin E2 in the context of viral infection. Our studies demonstrate that a viral miRNA mediates translational repression of multiple cellular genes by targeting mRNA 5'UTRs.
Grey
Finn
F
Vaccine and Gene Therapy Institute, Oregon Health & Science University, Portland, Oregon, United States of America. finn.grey@roslin.ed.ac.uk
Tirabassi
Rebecca
R
Meyers
Heather
H
Wu
Guanming
G
McWeeney
Shannon
S
Hook
Lauren
L
Nelson
Jay A
JA
D020121:5' Untranslated Regions / Q000235:genetics*; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000235:genetics*; D002478:Cells, Cultured; D016213:Cyclins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D003587:Cytomegalovirus / Q000502:physiology; D003586:Cytomegalovirus Infections / Q000235:genetics / Q000821:virology; D015536:Down-Regulation; D005786:Gene Expression Regulation; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics*; D034743:RNA-Induced Silencing Complex / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014779:Virus Replication / Q000235:genetics*
D020121:5' Untranslated Regions; C503688:CCNE2 protein, human; D018797:Cell Cycle Proteins; D016213:Cyclins; D035683:MicroRNAs; D012333:RNA, Messenger; D012367:RNA, Viral; D034743:RNA-Induced Silencing Complex; D008156:Luciferases