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

<h4>Background</h4>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 main...

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Autores principales:	Zhang-Zhou Shen, Xing Pan, Ling-Feng Miao, Han-Qing Ye, Stéphane Chavanas, Christian Davrinche, Michael McVoy, Min-Hua Luo
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e8b24b9da1bb4d6281887e493409ffac
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spelling	oai:doaj.org-article:e8b24b9da1bb4d6281887e493409ffac2021-11-18T08:32:49ZComprehensive analysis of human cytomegalovirus microRNA expression during lytic and quiescent infection.1932-620310.1371/journal.pone.0088531https://doaj.org/article/e8b24b9da1bb4d6281887e493409ffac2014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24533100/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>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.<h4>Methods</h4>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.<h4>Results</h4>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.<h4>Conclusions</h4>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.Zhang-Zhou ShenXing PanLing-Feng MiaoHan-Qing YeStéphane ChavanasChristian DavrincheMichael McVoyMin-Hua LuoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 2, p e88531 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Zhang-Zhou Shen Xing Pan Ling-Feng Miao Han-Qing Ye Stéphane Chavanas Christian Davrinche Michael McVoy Min-Hua Luo Comprehensive analysis of human cytomegalovirus microRNA expression during lytic and quiescent infection.
description	<h4>Background</h4>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.<h4>Methods</h4>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.<h4>Results</h4>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.<h4>Conclusions</h4>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.
format	article
author	Zhang-Zhou Shen Xing Pan Ling-Feng Miao Han-Qing Ye Stéphane Chavanas Christian Davrinche Michael McVoy Min-Hua Luo
author_facet	Zhang-Zhou Shen Xing Pan Ling-Feng Miao Han-Qing Ye Stéphane Chavanas Christian Davrinche Michael McVoy Min-Hua Luo
author_sort	Zhang-Zhou Shen
title	Comprehensive analysis of human cytomegalovirus microRNA expression during lytic and quiescent infection.
title_short	Comprehensive analysis of human cytomegalovirus microRNA expression during lytic and quiescent infection.
title_full	Comprehensive analysis of human cytomegalovirus microRNA expression during lytic and quiescent infection.
title_fullStr	Comprehensive analysis of human cytomegalovirus microRNA expression during lytic and quiescent infection.
title_full_unstemmed	Comprehensive analysis of human cytomegalovirus microRNA expression during lytic and quiescent infection.
title_sort	comprehensive analysis of human cytomegalovirus microrna expression during lytic and quiescent infection.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/e8b24b9da1bb4d6281887e493409ffac
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Comprehensive analysis of human cytomegalovirus microRNA expression during lytic and quiescent infection.

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