+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Comprehensive analysis of human cytomegalovirus microRNA expression during lytic and quiescent infection



Comprehensive analysis of human cytomegalovirus microRNA expression during lytic and quiescent infection



Plos one 9(2): E88531



Human cytomegalovirus (HCMV) encodes microRNAs (miRNAs) that function as post-transcriptional regulators of gene expression during lytic infection in permissive cells. Some miRNAs have been shown to suppress virus replication, which could help HCMV to establish or maintain latent infection. However, HCMV miRNA expression has not been comprehensively examined and compared using cell culture systems representing permissive (lytic) and semi-permissive vs. non-permissive (latent-like) infection. Viral miRNAs levels and expression kinetics during HCMV infection were determined by miRNA-specific stem-loop RT-PCR. HCMV infected THP-1 (non-permissive), differentiated THP-1 (d-THP-1, semi-permissive) and human embryo lung fibroblasts (HELs, fully-permissive) were examined. The impact of selected miRNAs on HCMV infection (gene expression, genome replication and virus release) was determined by Western blotting, RT-PCR, qPCR, and plaque assay. Abundant expression of 15 HCMV miRNAs was observed during lytic infection in HELs; highest peak inductions (11- to 1502-fold) occurred at 48 hpi. In d-THP-1s, fourteen mRNAs were detected with moderate induction (3- to 288-fold), but kinetics of expression was generally delayed for 24 h relative to HELs. In contrast, only three miRNAs were induced to low levels (3- to 4-fold) during quiescent infection in THP-1s. Interestingly, miR-UL70-3p was poorly induced in HEL (1.5-fold), moderately in THP-1s (4-fold), and strongly (58-fold) in d-THP-1s, suggesting a potentially specific role for miR-UL70-3p in THP-1s and d-THP-1s. MiR-US33, -UL22A and -UL70 were further evaluated for their impact on HCMV replication in HELs. Ectopic expression of miR-UL22A and miR-UL70 did not affect HCMV replication in HELs, whereas miR-US33 inhibited HCMV replication and reduced levels of HCMV US29 mRNA, confirming that US29 is a target of miR-US33. Viral miRNA expression kinetics differs between permissive, semi-permissive and quiescent infections, and miR-US33 down-regulates HCMV replication. These results suggest that miR-US33 may function to impair entry into lytic replication and hence promote establishment of latency.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 052269801

Download citation: RISBibTeXText

PMID: 24533100

DOI: 10.1371/journal.pone.0088531


Related references

Human cytomegalovirus infection induces cellular thymidylate synthase gene expression in quiescent fibroblasts. Journal of General Virology 83(Pt 12): 2983-2993, 2002

Comprehensive analysis of microRNA expression in regionalized human neural progenitor cells reveals microRNA-10 as a caudalizing factor. Development 142(18): 3166-3177, 2015

Chromatin structure regulates human cytomegalovirus gene expression during latency, reactivation and lytic infection. Biochimica et Biophysica Acta 1799(3-4): 286-295, 2010

Simultaneous integration of microRNA and mRNA expression in the analysis of quiescent and activated human fibroblasts in the diseased pancreas. Pancreatology 15(3): S19-S20, 2015

Human cytomegalovirus latent infection alters the expression of cellular and viral microRNA. Gene 536(2): 272-278, 2014

Control of human cytomegalovirus gene expression by differential histone modifications during lytic and latent infection of a monocytic cell line. Gene 384: 120-128, 2006

Human cytomegalovirus IE72 protein interacts with the transcriptional repressor hDaxx to regulate LUNA gene expression during lytic infection. Journal of Virology 84(14): 7185-7194, 2010

Analysis of human cytomegalovirus-encoded microRNA activity during infection. Journal of Virology 83(20): 10684-10693, 2009

Signature microRNA expression profile of essential hypertension and its novel link to human cytomegalovirus infection. Circulation 124(2): 175-184, 2011

Human cytomegalovirus infection alters the expression of cellular microRNA species that affect its replication. Journal of Virology 82(18): 9065-9074, 2008

Early stage of cytomegalovirus infection suppresses host microRNA expression regulation in human fibroblasts. Cell Cycle 15(24): 3378-3389, 2016

Lytic infection of permissive cells with human cytomegalovirus is regulated by an intrinsic 'pre-immediate-early' repression of viral gene expression mediated by histone post-translational modification. Journal of General Virology 90(Pt 10): 2364-2374, 2009

Response to Letter Regarding Article, "Signature MicroRna Expression Profile of Essential Hypertension and Its Novel Link to Human Cytomegalovirus Infection". Circulation 125(5): e338-e339, 2012

Comprehensive mapping and analysis of Kaposi's sarcoma-associated herpesvirus 3' UTRs identify differential posttranscriptional control of gene expression in lytic versus latent infection. Journal of Virology 87(23): 12838-12849, 2013

Altered microRNA expression after infection with human cytomegalovirus leads to TIMP3 downregulation and increased shedding of metalloprotease substrates, including MICA. Journal of Immunology 193(3): 1344-1352, 2014