Faculty Opinions recommendation of An RNA Thermometer Activity of the West Nile Virus Genomic 3'-Terminal Stem-Loop Element Modulates Viral Replication Efficiency during Host Switching.

2020 ◽  
Author(s):  
Franz Narberhaus
Keyword(s):  
West Nile Virus ◽  
Viral Replication ◽  
Host Switching ◽  
The West ◽  
West Nile ◽  
Stem Loop ◽  
Replication Efficiency ◽  
Loop Element ◽  
Rna Thermometer ◽  
Viral Replication Efficiency

scholarly journals An RNA Thermometer Activity of the West Nile Virus Genomic 3′-Terminal Stem-Loop Element Modulates Viral Replication Efficiency during Host Switching

Viruses ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 104 ◽  
Author(s):  
Alexandra Meyer ◽  
Marie Freier ◽  
Tobias Schmidt ◽  
Katja Rostowski ◽  
Juliane Zwoch ◽  
...  
Keyword(s):  
Viral Replication ◽  
Rna Binding ◽  
Rna Replication ◽  
Viral Rna ◽  
Cell Protein ◽  
Host Switching ◽  
West Nile ◽  
Stem Loop ◽  
Mosquito Cells ◽  
Rna Thermometer

The 3′-terminal stem-loop (3′SL) of the RNA genome of the flavivirus West Nile (WNV) harbors, in its stem, one of the sequence elements that are required for genome cyclization. As cyclization is a prerequisite for the initiation of viral replication, the 3′SL was proposed to act as a replication silencer. The lower part of the 3′SL is metastable and confers a structural flexibility that may regulate the switch from the linear to the circular conformation of the viral RNA. In the human system, we previously demonstrated that a cellular RNA-binding protein, AUF1 p45, destabilizes the 3′SL, exposes the cyclization sequence, and thus promotes flaviviral genome cyclization and RNA replication. By investigating mutant RNAs with increased 3′SL stabilities, we showed the specific conformation of the metastable element to be a critical determinant of the helix-destabilizing RNA chaperone activity of AUF1 p45 and of the precision and efficiency of the AUF1 p45-supported initiation of RNA replication. Studies of stability-increasing mutant WNV replicons in human and mosquito cells revealed that the cultivation temperature considerably affected the replication efficiencies of the viral RNA variants and demonstrated the silencing effect of the 3′SL to be temperature dependent. Furthermore, we identified and characterized mosquito proteins displaying similar activities as AUF1 p45. However, as the RNA remodeling activities of the mosquito proteins were found to be considerably lower than those of the human protein, a potential cell protein-mediated destabilization of the 3′SL was suggested to be less efficient in mosquito cells. In summary, our data support a model in which the 3′SL acts as an RNA thermometer that modulates flavivirus replication during host switching.

scholarly journals Cell Proteins TIA-1 and TIAR Interact with the 3′ Stem-Loop of the West Nile Virus Complementary Minus-Strand RNA and Facilitate Virus Replication

2002 ◽  
Vol 76 (23) ◽  
pp. 11989-12000 ◽  
Author(s):  
W. Li ◽  
Y. Li ◽  
N. Kedersha ◽  
P. Anderson ◽  
M. Emara ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Rna Binding ◽  
Peptide Sequencing ◽  
Viral Rna ◽  
Cell Protein ◽  
Affinity Column ◽  
Rna Recognition Motif ◽  
The West ◽  
West Nile ◽  
Stem Loop

ABSTRACT It was reported previously that four baby hamster kidney (BHK) proteins with molecular masses of 108, 60, 50, and 42 kDa bind specifically to the 3′-terminal stem-loop of the West Nile virus minus-stand RNA [WNV 3′(−) SL RNA] (P. Y. Shi, W. Li, and M. A. Brinton, J. Virol. 70:6278-6287, 1996). In this study, p42 was purified using an RNA affinity column and identified as TIAR by peptide sequencing. A 42-kDa UV-cross-linked viral RNA-cell protein complex formed in BHK cytoplasmic extracts incubated with the WNV 3′(−) SL RNA was immunoprecipitated by anti-TIAR antibody. Both TIAR and the closely related protein TIA-1 are members of the RNA recognition motif (RRM) family of RNA binding proteins. TIA-1 also binds to the WNV 3′(−) SL RNA. The specificity of these viral RNA-cell protein interactions was demonstrated using recombinant proteins in competition gel mobility shift assays. The binding site for the WNV 3′(−) SL RNA was mapped to RRM2 on both TIAR and TIA-1. However, the dissociation constant (Kd ) for the interaction between TIAR RRM2 and the WNV 3′(−) SL RNA was 1.5 × 10−8, while that for TIA-1 RRM2 was 1.12 × 10−7. WNV growth was less efficient in murine TIAR knockout cell lines than in control cells. This effect was not observed for two other types of RNA viruses or two types of DNA viruses. Reconstitution of the TIAR knockout cells with TIAR increased the efficiency of WNV growth, but neither the level of TIAR nor WNV replication was as high as in control cells. These data suggest a functional role for TIAR and possibly also for TIA-1 during WNV replication.

scholarly journals Genetic Interactions among the West Nile Virus Methyltransferase, the RNA-Dependent RNA Polymerase, and the 5′ Stem-Loop of Genomic RNA

2008 ◽  
Vol 82 (14) ◽  
pp. 7047-7058 ◽  
Author(s):  
Bo Zhang ◽  
Hongping Dong ◽  
Yangsheng Zhou ◽  
Pei-Yong Shi
Keyword(s):  
West Nile Virus ◽  
Rna Polymerase ◽  
Viral Replication ◽  
Genetic Interactions ◽  
Genome Length ◽  
Genomic Rna ◽  
Rna Dependent Rna Polymerase ◽  
West Nile ◽  
Stem Loop ◽  
Compensatory Mutations

ABSTRACT Flavivirus methyltransferase catalyzes both guanine N7 and ribose 2′-OH methylations of the viral RNA cap (GpppA-RNA→m7GpppAm-RNA). The methyltransferase is physically linked to an RNA-dependent RNA polymerase (RdRp) in the flaviviral NS5 protein. Here, we report genetic interactions of West Nile virus (WNV) methyltransferase with the RdRp and the 5′-terminal stem-loop of viral genomic RNA. Genome-length RNAs, containing amino acid substitutions of D146 (a residue essential for both cap methylations) in the methyltransferase, were transfected into BHK-21 cells. Among the four mutant RNAs (D146L, D146P, D146R, and D146S), only D146S RNA generated viruses in transfected cells. Sequencing of the recovered viruses revealed that, besides the D146S change in the methyltransferase, two classes of compensatory mutations had reproducibly emerged. Class 1 mutations were located in the 5′-terminal stem-loop of the genomic RNA (a G35U substitution or U38 insertion). Class 2 mutations resided in NS5 (K61Q in methyltransferase and W751R in RdRp). Mutagenesis analysis, using a genome-length RNA and a replicon of WNV, demonstrated that the D146S substitution alone was lethal for viral replication; however, the compensatory mutations rescued replication, with the highest rescuing efficiency occurring when both classes of mutations were present. Biochemical analysis showed that a low level of N7 methylation of the D146S methyltransferase is essential for the recovery of adaptive viruses. The methyltransferase K61Q mutation facilitates viral replication through improved N7 methylation activity. The RdRp W751R mutation improves viral replication through an enhanced polymerase activity. Our results have clearly established genetic interactions among flaviviral methyltransferase, RdRp, and the 5′ stem-loop of the genomic RNA.

scholarly journals Arginine methylation enhances the RNA chaperone activity of the West Nile virus host factor AUF1 p45

RNA ◽  
2016 ◽  
Vol 22 (10) ◽  
pp. 1574-1591 ◽  
Author(s):  
Susann Friedrich ◽  
Tobias Schmidt ◽  
Angelika Schierhorn ◽  
Hauke Lilie ◽  
Grit Szczepankiewicz ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Arginine Methylation ◽  
Host Factor ◽  
Chaperone Activity ◽  
The West ◽  
West Nile ◽  
Rna Chaperone

Polymorphic microsatellite loci from the West Nile virus vector Culex tarsalis

2006 ◽  
Vol 6 (3) ◽  
pp. 680-682 ◽  
Author(s):  
JASON L. RASGON ◽  
MEERA VENKATESAN ◽  
CATHERINE J. WESTBROOK ◽  
MARY CLAIRE HAUER
Keyword(s):  
West Nile Virus ◽  
Microsatellite Loci ◽  
Virus Vector ◽  
Culex Tarsalis ◽  
The West ◽  
West Nile ◽  
Polymorphic Microsatellite
Sign in / Sign up

Export Citation Format

Share Document