scholarly journals The host antiviral ribonuclease L protein supports Zika virus replication factory formation to enhance infectious virus production

2019 ◽  
Author(s):  
Jillian N Whelan ◽  
Joshua Hatterschide ◽  
David M. Renner ◽  
Beihua Dong ◽  
Robert H Silverman ◽  
...  
Keyword(s):  
Zika Virus ◽  
Virus Production ◽  
Rnase L ◽  
Microtubule Stabilization ◽  
Replication Factory ◽  
Inactive Form ◽  
Antiviral Function ◽  
Cytoskeleton Rearrangements ◽  
Ribonuclease L

SummaryThe flavivirus Zika virus (ZIKV) activates ribonuclease L (RNase L) catalytic antiviral function during infection, yet deletion of RNase L decreases ZIKV production, suggesting a proviral role of RNase L. In this study, we reveal that latent RNase L supports ZIKV replication factory (RF) assembly. Deletion of RNase L induced broader cellular distribution of ZIKV dsRNA and NS3 compared with densely concentrated RFs detected in WT cells. An inactive form of RNase L was sufficient to contain ZIKV genome and dsRNA within a smaller area, which increased levels of viral RNA within RFs as well as infectious ZIKV released from the cell. We used a microtubule stabilization drug to demonstrate that RNase L deletion impaired the cytoskeleton rearrangements that are required for proper generation of RFs. During infection with dengue or West Nile Kunjin viruses, RNase L decreased virus production, suggesting that RNase L proviral function is specific to ZIKV.

scholarly journals Zika virus employs the host antiviral RNase L protein to support replication factory assembly

2021 ◽  
Vol 118 (22) ◽  
pp. e2101713118
Author(s):  
Jillian N. Whelan ◽  
Nicholas A. Parenti ◽  
Joshua Hatterschide ◽  
David M. Renner ◽  
Yize Li ◽  
...  
Keyword(s):  
Zika Virus ◽  
Virus Production ◽  
Rnase L ◽  
L Protein ◽  
Microtubule Stabilization ◽  
Replication Factory ◽  
Inactive Form ◽  
Route Of Transmission ◽  
Cytoskeleton Rearrangements ◽  
Ns3 Protease

Infection with the flavivirus Zika virus (ZIKV) can result in tissue tropism, disease outcome, and route of transmission distinct from those of other flaviviruses; therefore, we aimed to identify host machinery that exclusively promotes the ZIKV replication cycle, which can inform on differences at the organismal level. We previously reported that deletion of the host antiviral ribonuclease L (RNase L) protein decreases ZIKV production. Canonical RNase L catalytic activity typically restricts viral infection, including that of the flavivirus dengue virus (DENV), suggesting an unconventional, proviral RNase L function during ZIKV infection. In this study, we reveal that an inactive form of RNase L supports assembly of ZIKV replication factories (RFs) to enhance infectious virus production. Compared with the densely concentrated ZIKV RFs generated with RNase L present, deletion of RNase L induced broader subcellular distribution of ZIKV replication intermediate double-stranded RNA (dsRNA) and NS3 protease, two constituents of ZIKV RFs. An inactive form of RNase L was sufficient to contain ZIKV genome and dsRNA within a smaller RF area, which subsequently increased infectious ZIKV release from the cell. Inactive RNase L can interact with cytoskeleton, and flaviviruses remodel cytoskeleton to construct RFs. Thus, we used the microtubule-stabilization drug paclitaxel to demonstrate that ZIKV repurposes RNase L to facilitate the cytoskeleton rearrangements required for proper generation of RFs. During infection with flaviviruses DENV or West Nile Kunjin virus, inactive RNase L did not improve virus production, suggesting that a proviral RNase L role is not a general feature of all flavivirus infections.

scholarly journals Zika Virus Production Is Resistant to RNase L Antiviral Activity

2019 ◽  
Vol 93 (16) ◽  
Author(s):  
Jillian N. Whelan ◽  
Yize Li ◽  
Robert H. Silverman ◽  
Susan R. Weiss
Keyword(s):  
Antiviral Activity ◽  
Zika Virus ◽  
Viral Genome ◽  
Molecular Mechanisms ◽  
Virus Production ◽  
Type I ◽  
Rnase L ◽  
Oligoadenylate Synthetase ◽  
Zikv Infection ◽  
Initial Report

SUMMARYThere is currently no knowledge of how the emerging human pathogen Zika virus (ZIKV) interacts with the antiviral endoribonuclease L (RNase L) pathway during infection. Since activation of RNase L during infection typically limits virus production dramatically, we used CRISPR-Cas9 gene editing technology to knockout (KO) targeted host genes involved in the RNase L pathway to evaluate the effects of RNase L on ZIKV infection in human A549 cells. RNase L was activated in response to ZIKV infection, which degraded ZIKV genomic RNA. Surprisingly, despite viral genome reduction, RNase L activity did not reduce ZIKV infectious titers. In contrast, both the flavivirus dengue virus and the alphavirus Sindbis virus replicated to significantly higher titers in RNase L KO cells compared to wild-type (WT) cells. Using MAVS/RNase L double KO cells, we demonstrated that the absence of increased ZIKV production in RNase L KO cells was not due to compensation by enhanced type I interferon transcripts to thus inhibit virus production. Finally, when synthetic double-stranded RNA was detected by OAS3 to induce RNase L antiviral activity prior to ZIKV infection, we observed reduced ZIKV replication factory formation, as well as a 42-fold reduction in virus yield in WT but not RNase L KO cells. This study proposes that ZIKV evades RNase L antiviral activity by generating a viral genome reservoir protected from RNase L cleavage during early infection, allowing for sufficient virus production before RNase L activation is detectable.IMPORTANCEWith the onset of the 2015 ZIKV outbreak, ZIKV pathogenesis has been of extreme global public health interest, and a better understanding of interactions with the host would provide insight into molecular mechanisms driving the severe neurological outcomes of ZIKV disease. Here is the initial report on the relationship between ZIKV and the host oligoadenylate synthetase-RNase L (OAS-RNase L) system, a potent antiviral pathway effective at restricting replication of diverse viruses. Our study elucidated a unique mechanism whereby ZIKV production is impervious to antiviral RNase L activity, through a mechanism of viral RNA protection that is not mimicked during infection with numerous other RNase L-activating viruses, thus identifying a distinct replication strategy potentially important for ZIKV pathogenesis.

scholarly journals Chromosomal Lesion Suppression and Removal in Escherichia coli via Linear DNA Degradation

Genetics ◽  
2003 ◽  
Vol 163 (4) ◽  
pp. 1255-1271 ◽  
Author(s):  
Anabel Miranda ◽  
Andrei Kuzminov
Keyword(s):  
Dna Degradation ◽  
Dna Helicase ◽  
Structural Element ◽  
E Coli ◽  
Linear Dna ◽  
Replication Factory ◽  
Low Copy Number ◽  
Chromosomal Lesion ◽  
Recbcd Enzyme

Abstract RecBCD is a DNA helicase/exonuclease implicated in degradation of foreign linear DNA and in RecA-dependent recombinational repair of chromosomal lesions in E. coli. The low viability of recA recBC mutants vs. recA mutants indicates the existence of RecA-independent roles for RecBCD. To distinguish among possible RecA-independent roles of the RecBCD enzyme in replication, repair, and DNA degradation, we introduced wild-type and mutant combinations of the recBCD chromosomal region on a low-copy-number plasmid into a ΔrecA ΔrecBCD mutant and determined the viability of resulting strains. Our results argue against ideas that RecBCD is a structural element in the replication factory or is involved in RecA-independent repair of chromosomal lesions. We found that RecBCD-catalyzed DNA degradation is the only activity important for the recA-independent viability, suggesting that degradation of linear tails of σ-replicating chromosomes could be one of the RecBCD’s roles. However, since the weaker DNA degradation capacity due a combination of the RecBC helicase and ssDNA-specific exonucleases restores viability of the ΔrecA ΔrecBCD mutant to a significant extent, we favor suppression of chromosomal lesions via linear DNA degradation at reversed replication forks as the major RecA-independent role of the RecBCD enzyme.

The Role of Predation in Determining Traits of Aedes aegypti (Diptera: Culicidae) and Infection With Zika Virus

2021 ◽  
Author(s):  
Shawna Bellamy ◽  
Barry W Alto
Keyword(s):  
Aedes Aegypti ◽  
Zika Virus ◽  
Life History Traits ◽  
Immature Stages ◽  
Direct Effects ◽  
Infected Blood ◽  
Predator Prey ◽  
Susceptibility To Infection ◽  
Density Reduction

Abstract Non-lethal predator-prey interactions during the immature stages can cause significant changes to mosquito life history traits and their ability to transmit pathogens as adults. Treatment manipulations using mosquitoes Aedes aegypti (L.) and Toxoryhnchites rutilus (Coquillett) were performed during the immature stages to explore the potential impacts of non-lethal interactions on adult susceptibility to infection, disseminated infection and saliva infection of Ae. aegypti following ingestion of Zika virus-infected blood. Treatments inducing density reduction resulted in reduced development time and survivorship to adulthood. However, effects of treatment did not alter infection, dissemination, or saliva infection. These observations indicate that, while non-lethal predation may impact some traits that influence population dynamics and transmission of pathogens, there were no direct effects on mosquito-arbovirus interactions.

scholarly journals The possible role of cross-reactive dengue virus antibodies in Zika virus pathogenesis

2019 ◽  
Vol 15 (4) ◽  
pp. e1007640 ◽  
Author(s):  
Thomas Langerak ◽  
Noreen Mumtaz ◽  
Vera I. Tolk ◽  
Eric C. M. van Gorp ◽  
Byron E. Martina ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Zika Virus

Investigating the Potential Role of North American Animals as Hosts for Zika Virus

2017 ◽  
Vol 17 (3) ◽  
pp. 161-164 ◽  
Author(s):  
Izabela K. Ragan ◽  
Emily L. Blizzard ◽  
Paul Gordy ◽  
Richard A. Bowen
Keyword(s):  
North American ◽  
Zika Virus ◽  
Potential Role

scholarly journals Dysregulation of Antiviral Function of CD8+T Cells in the Chronic Obstructive Pulmonary Disease Lung. Role of the PD-1–PD-L1 Axis

2016 ◽  
Vol 193 (6) ◽  
pp. 642-651 ◽  
Author(s):  
Richard T. McKendry ◽  
C. Mirella Spalluto ◽  
Hannah Burke ◽  
Ben Nicholas ◽  
Doriana Cellura ◽  
...  
Keyword(s):  
Chronic Obstructive Pulmonary Disease ◽  
T Cells ◽  
Pulmonary Disease ◽  
Cd8 T Cells ◽  
Chronic Obstructive ◽  
Obstructive Pulmonary Disease ◽  
Antiviral Function

scholarly journals Overexpression of a Neural-specific Rho Family GTPase, cRac1B, Selectively Induces Enhanced Neuritogenesis and Neurite Branching in Primary Neurons

1998 ◽  
Vol 142 (3) ◽  
pp. 815-825 ◽  
Author(s):  
Chiara Albertinazzi ◽  
Daniela Gilardelli ◽  
Simona Paris ◽  
Renato Longhi ◽  
Ivan de Curtis
Keyword(s):  
Neural Development ◽  
Neuronal Morphology ◽  
Primary Neurons ◽  
Actin Organization ◽  
Inactive Form ◽  
Rho Family Gtpases ◽  
Neurite Formation ◽  
New Gene ◽  
Rho Family

Rho family GTPases have been implicated in cytoskeletal reorganization during neuritogenesis. We have recently identified a new gene of this family, cRac1B, specifically expressed in the chicken developing nervous system. This GTPase was overexpressed in primary neurons to study the role of cRac1B in the development of the neuronal phenotype. Overexpression of cRac1B induced an increment in the number of neurites per neuron, and dramatically increased neurite branching, whereas overexpression of the highly related and ubiquitous cRac1A GTPase did not evidently affect neuronal morphology. Furthermore, expression of an inactive form of cRac1B strikingly inhibited neurite formation. The specificity of cRac1B action observed in neurons was not observed in fibroblasts, where both GTPases produced similar effects on cell morphology and actin organization, indicating the existence of a cell type-dependent specificity of cRac1B function. Molecular dissection of cRac1B function by analysis of the effects of chimeric cRac1A/cRac1B proteins showed that the COOH-terminal portion of cRac1B is essential to induce increased neuritogenesis and neurite branching. Considering the distinctive regulation of cRac1B expression during neural development, our data strongly support an important role of cRac1B during neuritogenesis, and they uncover new mechanisms underlying the functional specificity of distinct Rho family GTPases.

scholarly journals A CYTOCHEMICAL DESCRIPTION OF THE MULTIPLICATION OF MENGOVIRUS IN L-929 CELLS

1962 ◽  
Vol 12 (1) ◽  
pp. 1-15 ◽  
Author(s):  
Richard M. Franklin
Keyword(s):  
Phase Contrast ◽  
Basic Protein ◽  
Virus Production ◽  
Cytological Change ◽  
Virus Particles ◽  
Contrast Microscopy ◽  
Infected Cells ◽  
Cytological Changes ◽  
Perinuclear Area

A correlation of cytochemical changes with virus production has been studied in L cells infected with Mengovirus. After a latent period of about 2 hours, virus was produced rapidly, reaching maximum titers of up to 12,000 particles per cell in 6 to 8 hours. The earliest cytological change was in the nucleus and consisted of a slight condensation of chromatin. There is no evidence, however, for the multiplication of either the viral RNA or protein in the nucleus. RNA, of high molecular weight, accumulated in the perinuclear area of the cytoplasm and was later found in inclusions. The perinuclear RNA was digestible with RNase and may be located in or on ribosomes. The inclusion RNA was resistant to RNase but could be removed by pepsin or potassium permanganate; it is probably in completed virus particles. Viral antigen was first observed in a perinuclear location and later in the above-mentioned inclusions. Although the viral protein contains appreciable amounts of arginine and lysine, it is not a basic protein of the histone type. Phase-contrast microscopy of living cells clearly demonstrated the role of the inclusions in release of virus from infected cells. A comparison is made between these cytological changes in Mengo-infected cells and those which have been found by other workers in polio-infected cells. There are many very similar changes.

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