Temporal Landscape of MicroRNA-Mediated Host-Virus Crosstalk during Productive Human Cytomegalovirus Infection

Kim, Sung‐Chul; Seo, Daekwan; Kim, Dongwoo; Hong, Yujin; Chang, Hyeshik; Baek, Daehyun; Kim, V. Narry; Lee, Sungwook; Ahn, Kwangseog

doi:10.1016/j.chom.2015.05.014

Cited by 59 publications

(86 citation statements)

References 45 publications

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“…Many results suggest that hcmv-miR-US25-1, or hcmv-miR-US25-1-5p, is involved in diverse process; not only cell cycle control, but also virus replication inhibition as well as apoptosis regulation [5,6,32]. It has been reported that hcmv-miR-US25-1-3p has the ability to downregulate CDK6 gene associated with suppress cell cycle progression [19,33]. Here, we identi ed highly HCMV-miRNA expression pattern upon reactivation from latency in vivo, especially for hcmv-miR-US25-1-3p in autoimmune disease patient, which provides a key foothold for the study of HCMV virology, even though HCMV-miRNAs' function remain to be study further in natural diseases such as in autoimmune disease.…”

Section: Discussionmentioning

confidence: 99%

Different Expression Pattern of Human Cytomegalovirus-Encoded microRNAs in Circulation from Virus Latency to Reactivation

Zhou

Wang

Ding

et al. 2020

Preprint

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Background: Human cytomegalovirus (HCMV) is a beta-hersvirinae that has a high latent infection rate worldwide and can cause serious consequences in immunocompromised patients when reactivation; however, the mechanism of how HCMV convert from latent to reactivation has rarely been investigated. In the present study, we aimed to perform a comprehensive analysis of the HCMV-encoded microRNA (miRNA) profile in serum of patients upon HCMV reactivation from latency and to further evaluate its clinical significance for the disease monitoring and preventing usefulness.Methods: Serum samples from 60 viremia patients and 60 age-gender matched controls were enrolled in this study for screening and validation of different expression of HCMV miRNAs. Serum concentrations of 22 known HCMV miRNAs were determined by a hydrolysis probe-based stem-loop quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay. HCMV DNA was measured by quantitative real-time PCR (qPCR) with the whole blood sample. Serum HCMV IgG and IgM were assessed using enzyme linked immunosorbent assay (ELISA). Another 47 samples from 5 patients at different time points were collected to evaluate the monitoring effectiveness and disease prediction ability of differential expression HCMV-miRNAs during the antiviral treatment. Results: The RT-qPCR analysis revealed that the serum levels of 16 of the 22 examined HCMV miRNAs were elevated in HCMV viremia patients compared with controls, and a profile of 8 HCMV miRNAs including hcmv-miR-US25-2-3p, hcmv-miR-US4-5p, hcmv-miR-US25-2-5p, hcmv-miR-US25-1-3p, hcmv-miR-US25-1, hcmv-miR-UL36, hcmv-miR-UL148D, hcmv-miR-US29-3p were markedly elevated (fold change > 2, P < 0.01). Receiver operating characteristic curve (ROC) analysis were performed on the selected HCMV-miRNAs in all of the patients and controls that enrolled in this study, and which ranged from 0.74 to 0.79 in the autoimmune patients. In addition, hcmv-miR-US25-1-3p levels were significantly correlated with HCMV DNA copies (r = 0.297，P = 0.022), and were obviously higher in the reactivation set than the latency set in the autoimmune patients, which could be a predictor for the monitoring of the antiviral treatment.Conclusions: HCMV miRNAs profile showed markedly shift-switch from latency to reactivation in circulation from HCMV infected patients and hcmv-miR-US25-1-3p may be served as a predictor for the switch upon reactivation from latency in patients suffered with autoimmune diseases.

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Section: Discussionmentioning

confidence: 99%

Different Expression Pattern of Human Cytomegalovirus-Encoded microRNAs in Circulation from Virus Latency to Reactivation

Zhou

Wang

Ding

et al. 2020

Preprint

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“…Thus, we investigated whether FOXO3a could also be a target of HCMV miRNAs. Biochemical and bioinformatic analysis (16) suggested that FOXO3a was a target of HCMV miR-US5-1 and miR-UL112-3p. We cloned the 3' UTR of FOXO3a or 3'…”

Section: Hcmv Mir-us5-1 and Mir-ul112 Cooperatively Target Foxo3amentioning

confidence: 99%

“…Additionally, miR-UL148D protects cells from apoptosis induced by overexpression of IEX-1 (15). Finally, Kim et al reported FAS as a target of miR-UL36-3p, miR-US5-1, and miR-US5-2-3p, Fas associated protein with death domain (FADD) as a target of miR-US5-1, Caspase 3 as a target of miR-US25-2-3p, miR-112-5p, and miR-UL22A-5p, Caspase 2 as a target of miR-US4-5p and Capase 7 as a target of miR-UL22A-3p and miR-US4-3p (16).…”

Section: Introductionmentioning

confidence: 99%

Human cytomegalovirus UL7, miR-US5-1 and miR-UL112-3p inactivation of FOXO3a protects CD34⁺hematopoietic progenitor cells from apoptosis

Hancock

Crawford

Perez

et al. 2020

Preprint

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Human cytomegalovirus (HCMV) infection of myeloid-lineage cells, such as CD34+ hematopoietic progenitor cells (HPCs) or monocytes results in the upregulation of anti-apoptotic cellular proteins that protect the newly infected cells from programmed cell death. The mechanisms used by HCMV to regulate pro-apoptotic cellular proteins upon infection of CD34+ HPCs has not been fully explored. Here we show that HCMV utilizes pUL7, a secreted protein that signals through the FLT3 receptor, and miR-US5-1 and miR-UL112-3p to reduce the abundance and activity of the pro-apoptotic transcription factor FOXO3a at early times after infection of CD34+ HPCs. Regulation of FOXO3a by pUL7, miR-US5-1 and miR-UL112 results in reduced expression of the pro-apoptotic BCL2L11 transcript and protection of CD34+ HPCs from virus-induced apoptosis. These data highlight the importance of both viral proteins and miRNAs in protecting CD34+ HPCs from apoptosis at early times post-infection, allowing for the establishment of latency and maintenance of viral genome-containing cells.ImportanceHuman cytomegalovirus (HCMV) causes serious disease in immunocompromised individuals and is a significant problem during transplantation. The virus can establish a latent infection in CD34+ hematopoietic progenitor cells (HPCs) and periodically reactivate to cause disease in the absence of an intact immune system. What viral gene products are required for successful establishment of latency are still not fully understood. Here we show that both a viral protein and viral miRNAs are required to prevent apoptosis after infection of CD34+ HPCs. HCMV pUL7 and miRNAs miR-US5-1 and miR-UL112-3p act to limit the expression and activation of the transcription factor FOXO3a which in turn reduces expression of pro-apoptotic gene BCL2L11 and prevents virus-induced apoptosis in CD34+ HPCs.

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“…The final complex was diluted in Storage buffer with 50% glycerol, snapfrozen in liquid nitrogen and stored at -80 °C. Mutant Cas1-Cas2 complexes were cloned by site-directed mutagenesis as described previously (Kim et al, 2015) and purified as described above. Primers for sub-cloning are listed in Table S1.…”

Section: Protein Preparationmentioning

confidence: 99%

Selective Prespacer Processing Ensures Precise CRISPR-Cas Adaptation

Loeff

Colombo

et al. 2019

Preprint

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CRISPR-Cas immunity protects prokaryotes against foreign genetic elements. CRISPR-Cas uses the highly conserved Cas1-Cas2 complex to establish inheritable memory (spacers). It remains elusive how Cas1-Cas2 acquires spacers from cellular DNA fragments (prespacers) and how it integrates them into the CRISPR array in the correct orientation. By using the high spatiotemporal resolution of single-molecule fluorescence, we reveal that Cas1-Cas2 obtains prespacers in various forms including single-stranded DNA and partial duplexes by selecting them in the DNA-length and PAM-dependent manner. Furthermore, we identify DnaQ exonucleases as enzymes that can mature the Cas1-Cas2-loaded precursor prespacers into an integration-competent size. Cas1-Cas2 protects the PAM sequence from maturation, which results in the production of asymmetrically trimmed prespacers and subsequent spacer integration in the correct orientation. This kinetic coordination in prespacer selection and PAM trimming provides comprehensive understanding of the mechanisms that underlie the integration of functional spacers in the CRISPR array.

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Temporal Landscape of MicroRNA-Mediated Host-Virus Crosstalk during Productive Human Cytomegalovirus Infection

Cited by 59 publications

References 45 publications

Different Expression Pattern of Human Cytomegalovirus-Encoded microRNAs in Circulation from Virus Latency to Reactivation

Different Expression Pattern of Human Cytomegalovirus-Encoded microRNAs in Circulation from Virus Latency to Reactivation

Human cytomegalovirus UL7, miR-US5-1 and miR-UL112-3p inactivation of FOXO3a protects CD34⁺hematopoietic progenitor cells from apoptosis

Selective Prespacer Processing Ensures Precise CRISPR-Cas Adaptation

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Temporal Landscape of MicroRNA-Mediated Host-Virus Crosstalk during Productive Human Cytomegalovirus Infection

Cited by 59 publications

References 45 publications

Different Expression Pattern of Human Cytomegalovirus-Encoded microRNAs in Circulation from Virus Latency to Reactivation

Different Expression Pattern of Human Cytomegalovirus-Encoded microRNAs in Circulation from Virus Latency to Reactivation

Human cytomegalovirus UL7, miR-US5-1 and miR-UL112-3p inactivation of FOXO3a protects CD34+hematopoietic progenitor cells from apoptosis

Selective Prespacer Processing Ensures Precise CRISPR-Cas Adaptation

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Human cytomegalovirus UL7, miR-US5-1 and miR-UL112-3p inactivation of FOXO3a protects CD34⁺hematopoietic progenitor cells from apoptosis