Comprehensive analysis of human cytomegalovirus microRNA expression during lytic and quiescent infection.
Comprehensive analysis of human cytomegalovirus microRNA expression during lytic and quiescent infection.
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裂解感染和静止感染期间人巨细胞病毒 MicroRNA 表达的综合分析
DOI：
10.1371/journal.pone.0088531
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Luo MH
中科院分区：
文献类型：
--
作者：
Shen ZZ;Pan X;Miao LF;Ye HQ;Chavanas S;Davrinche C;McVoy M;Luo MH
人巨细胞病毒(HCMV)编码的microRNAs(MiRNAs)在允许细胞的裂解感染过程中作为基因表达的转录后调节因子发挥作用。一些miRNAs已被证明可以抑制病毒复制，这可能有助于建立或维持HCMV的潜伏感染。然而，利用代表许可(溶解)和半许可与非许可(潜伏性)感染的细胞培养系统，还没有全面地检查和比较HCMV miRNA的表达。用miRNA特异的茎环RT-PCR检测病毒在HCMV感染过程中的miRNAs水平和表达动力学。检测人巨细胞病毒感染THP-1(不允许)、分化THP-1(d-THP-1，半允许)和人胚肺成纤维细胞(HELs，完全允许)。通过Western blotting、RT-PCR、qPCR和空斑实验确定所选miRNAs对HCMV感染(基因表达、基因组复制和病毒释放)的影响。在HELs的裂解感染过程中，观察到15个HCMV miRNAs的大量表达；高峰诱导(11-1502倍)出现在48HPI。在d-THP-1s中，检测到14个中等诱导的mRNAs(3-288倍)，但表达动力学通常比HELs延迟24小时。相反，在THP-1s静止感染期间，只有3个miRNAs被诱导到低水平(3-4倍)。有趣的是，miR-UL70-3p在HEL中诱导较弱(1.5倍)，在THP-1s中诱导中等(4倍)，在d-THP-1s中诱导强烈(58倍)，提示miR-UL70-3p在THP-1s和d-THP-1s中具有潜在的特异性作用。进一步评估MIR-US33、-UL22A和-UL70对人巨细胞病毒复制的影响。MiR-UL22A和miR-UL70的异位表达不影响HCMV在HELs中的复制，而miR-US33抑制了HCMV的复制并降低了HCMV US29的mRNA水平，证实了US29是miR-US33的靶点。病毒miRNA在允许、半允许和静止感染中的表达动力学不同，miR-US33下调HCMV的复制。这些结果表明，miR-US33可能会损害进入裂解复制，从而促进潜伏期的建立。
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.
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影响因子：
14.9
作者：
Chen C;Ridzon DA;Broomer AJ;Zhou Z;Lee DH;Nguyen JT;Barbisin M;Xu NL;Mahuvakar VR;Andersen MR;Lao KQ;Livak KJ;Guegler KJ
通讯作者：
Guegler KJ
卡波西肉瘤相关疱疹病毒编码的 miR-K12-7-5p 通过靶向病毒 ORF50/RTA 稳定潜伏状态
影响因子：
3.7
作者：
Lin X;Liang D;He Z;Deng Q;Robertson ES;Lan K
通讯作者：
Lan K
DOI：
10.1007/978-1-60761-533-0_6
发表时间：
2010-01-01
期刊：
LENTIVIRUS GENE ENGINEERING PROTOCOLS, SECOND EDITION
影响因子：
--
作者：
Michelini, Zuleika;Negri, Donatella;Cara, Andrea
通讯作者：
Cara, Andrea
爱泼斯坦 - 巴尔病毒编码的microRNA mir-bart2下调病毒DNA聚合酶BALF5。
影响因子：
14.9
作者：
Barth, Stephanie;Pfuhl, Thorsten;Mamiani, Alfredo;Ehses, Claudia;Roemer, Klaus;Kremmer, Elisabeth;Jaeker, Christoph;Hoeck, Julia;Meister, Gunter;Graesser, Friedrich A.
通讯作者：
Graesser, Friedrich A.
影响因子：
14.9
作者：
Griffiths-Jones S;Grocock RJ;van Dongen S;Bateman A;Enright AJ
通讯作者：
Enright AJ