Tick-Borne Encephalitis Virus Nonstructural Protein NS5 Induces RANTES Expression Dependent on the RNA-Dependent RNA Polymerase Activity

Zheng, Zifeng; Yang, Junyi; Jiang, Xuan; Liu, Yalan; Zhang, Xiaowei; Li, Mei; Zhang, Mudan; Fu, Ming; Hu, Kai; Wang, Hanzhong; Luo, Min‐Hua; Gong, Peng; Hu, Qinxue

doi:10.4049/jimmunol.1701507

Cited by 30 publications

(37 citation statements)

References 90 publications

(101 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…On one hand, this helix is critical to both the JEV-mode and DENV3-mode of interface interactions by contributing the F349 and R353 residues (Fig 2A and 2B; Fig 3B). On the other hand, it is at the central region of a long stretch of conserved sequences (residues 341-366 in DENV2 NS5) that may also be related to NS5 nuclear localization, nuclear export, and interaction with another important viral protein NS3 [19,30,31], emphasizing its possible importance when not participating in the intra-molecular MTase-RdRP interactions. We propose that the largely rotational movement of the MTase from the JEV-mode conformations to the DENV3-mode conformations may utilize this highly conserved helix as a guiding track (S1 Movie).…”

Section: Plos Pathogensmentioning

confidence: 99%

A conformation-based intra-molecular initiation factor identified in the flavivirus RNA-dependent RNA polymerase

Zhang

et al. 2020

PLoS Pathog

Self Cite

View full text Add to dashboard Cite

The flaviviruses pose serious threats to human health. Being a natural fusion of a methyltransferase (MTase) and an RNA-dependent RNA polymerase (RdRP), NS5 is the most conserved flavivirus protein and an important antiviral target. Previously reported NS5 structures represented by those from the Japanese encephalitis virus (JEV) and Dengue virus serotype 3 (DENV3) exhibit two apparently different global conformations, defining two sets of intra-molecular MTase-RdRP interactions. However, whether these NS5 conformations are conserved in flaviviruses and their specific functions remain elusive. Here we report two forms of DENV serotype 2 (DENV2) NS5 crystal structures representing two conformational states with defined analogies to the JEV-mode and DENV3-mode conformations, respectively, demonstrating the conservation of both conformation modes and providing clues for how different conformational states may be interconnected. Data from in vitro polymerase assays further demonstrate that perturbing the JEV-mode but not the DENV3-mode intramolecular interactions inhibits catalysis only at initiation, while the cell-based virological analysis suggests that both modes of interactions are important for virus proliferation. Our work highlights the role of MTase as a unique intra-molecular initiation factor specifically only through the JEV-mode conformation, providing an example of conformation-based crosstalk between naturally fused protein functional modules.

show abstract

Section: Plos Pathogensmentioning

confidence: 99%

A conformation-based intra-molecular initiation factor identified in the flavivirus RNA-dependent RNA polymerase

Zhang

et al. 2020

PLoS Pathog

Self Cite

View full text Add to dashboard Cite

show abstract

“…The siRNAs targeting vacuolar ATPase (VTPase, sc-42686), dynamin (sc-43736), Rab5 (sc-36344), and Rab7 (sc-29460), respectively, were purchased from Santa Cruz. T98G cells were transfected with the indicated siRNAs using a HiPerFect Transfection Reagent (Qiagen) according to the manufacturer's protocol (Zheng et al, 2018). Knockdown efficiency was measured by Western blot.…”

Section: Rna-mediated Interferencementioning

confidence: 99%

Characterization of Zika Virus Endocytic Pathways in Human Glioblastoma Cells

Zhang

et al. 2020

Front. Microbiol.

Self Cite

View full text Add to dashboard Cite

Zika virus (ZIKV) infections can cause microcephaly and neurological disorders. However, the early infection events of ZIKV in neural cells remain to be characterized. Here, by using a combination of pharmacological and molecular approaches and the human glioblastoma cell T98G as a model, we first observed that ZIKV infection was inhibited by chloroquine and NH 4 Cl, indicating a requirement of low intracellular pH. We further showed that dynamin is required as the ZIKV entry was affected by the specific inhibitor dynasore, small interfering RNA (siRNA) knockdown of dynamin, or by expressing the dominant-negative K44A mutant. Moreover, the ZIKV entry was significantly inhibited by chlorpromazine, pitstop2, or siRNA knockdown of clathrin heavy chain, indicating an involvement of clathrin-mediated endocytosis. In addition, genistein treatment, siRNA knockdown of caveolin-1, or overexpression of a dominantnegative caveolin mutant impacted the ZIKV entry, with ZIKV particles being observed to colocalize with caveolin-1, implying that caveola endocytosis can also be involved. Furthermore, we found that the endocytosis of ZIKV is dependent on membrane cholesterol, microtubules, and actin cytoskeleton. Importantly, ZIKV infection was inhibited by silencing of Rab5 and Rab7, while confocal microscopy showed that ZIKV particles localized in Rab5-and Rab7-postive endosomes. These results indicated that, after internalization, ZIKV likely moves to Rab5-positive early endosome and Rab7positive late endosomes before delivering its RNA into the cytoplasm. Taken together, our study, for the first time, described the early infection events of ZIKV in human glioblastoma cell T98G.

show abstract

“…During TBEV infection of human astrocytes, the RLRs RIG-I and MDA5, but not TLR3, sense TBEV NS5 and activate IRF-3 signaling, which leads to the production of CCL5 (58). A recent study analyzing RNA expression profiles of TBEV infected human neuro-glia cultures, i.e., neuron and astrocyte cocultures, showed that in astrocytes the transcripts of Ddx58, Oas2, IFN-β, Mx1, Trim5a and Rasd2 were even more abundantly induced than in neurons (57).…”

Section: Discussionmentioning

confidence: 99%

Sequential MAVS- and MyD88/TRIF-signaling triggers anti-viral responses of tick-borne encephalitis virus-infected murine astrocytes

Ghita

Breitkopf

Mulenge

et al. 2020

Preprint

View full text Add to dashboard Cite

AbstractTick-borne encephalitis virus (TBEV), a member of the Flaviviridae family, is typically transmitted upon tick bite and can cause meningitis and encephalitis in humans. In TBEV infected mice, mitochondrial antiviral signaling protein (MAVS), the downstream adaptor of retinoic acid inducible gene I-like receptor (RLR)-signaling, is needed to induce early type I interferon (IFN) responses and to confer protection. To identify the brain resident cell subset that produces protective IFN-β in TBEV infected mice, we isolated neurons, astrocytes and microglia and exposed these cells to TBEV in vitro. Under such conditions, neurons showed the highest percentage of infected cells, whereas astrocytes and microglia were infected to a lesser extent. In the supernatant (SN) of infected neurons, IFN-β was not detectable, while infected astrocytes showed very high and microglia low IFN-β production. Transcriptome analyses of astrocytes implied that MAVS-signaling was needed early after TBEV infection. Accordingly, MAVS-deficient astrocytes showed enhanced TBEV infection and significantly reduced early IFN-β responses. At later time points, moderate amounts of IFN-β were detected in the SN of infected MAVS-deficient astrocytes. Transcriptome analyses indicated that MAVS-deficiency negatively affected the induction of early anti-viral responses, which resulted in significantly increased TBEV replication. Treatment with MyD88 and TRIF inhibiting peptides reduced late IFN-β responses of TBEV infected WT astrocytes and entirely blocked IFN-β responses of infected MAVS-deficient astrocytes. Thus, upon TBEV exposure of brain-resident cells, astrocytes are important IFN-β producers that show biphasic IFN-β induction that initially depends on MAVS- and later on MyD88/TRIF-signaling.

show abstract

Tick-Borne Encephalitis Virus Nonstructural Protein NS5 Induces RANTES Expression Dependent on the RNA-Dependent RNA Polymerase Activity

Cited by 30 publications

References 90 publications

A conformation-based intra-molecular initiation factor identified in the flavivirus RNA-dependent RNA polymerase

A conformation-based intra-molecular initiation factor identified in the flavivirus RNA-dependent RNA polymerase

Characterization of Zika Virus Endocytic Pathways in Human Glioblastoma Cells

Sequential MAVS- and MyD88/TRIF-signaling triggers anti-viral responses of tick-borne encephalitis virus-infected murine astrocytes

Contact Info

Product

Resources

About