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RSV filament

Masfique Mehedi, Margery Smelkinson, Juraj Kabat, Sundar Ganesan, Peter L Collins, Ursula J Buchholz
Human respiratory syncytial virus (RSV) infection in human lung epithelial A549 cells induces filopodia, cellular protrusions consisting of F-actin, that extend to neighboring uninfected cells (Mehedi et al ., 2016). High-resolution imaging via stimulated emission depletion (STED) microscopy revealed filamentous RSV particles along these filopodia, suggesting that filopodia facilitate RSV cell-to-cell spread (Mehedi et al ., 2016). In this protocol, we describe how to fix, permeabilize, immunostain, and mount RSV-infected A549 cells for STED imaging...
September 5, 2017: Bio-protocol
Daryll Vanover, Daisy V Smith, Emmeline L Blanchard, Eric Alonas, Jonathan L Kirschman, Aaron W Lifland, Chiara Zurla, Philip J Santangelo
The human respiratory syncytial virus G protein plays an important role in the entry and assembly of filamentous virions. Here, we report the use of fluorescently labeled soybean agglutinin to selectively label the respiratory syncytial virus G protein in living cells without disrupting respiratory syncytial virus infectivity or filament formation and allowing for interrogations of respiratory syncytial virus virion assembly. Using this approach, we discovered that plasma membrane-bound respiratory syncytial virus G rapidly recycles from the membrane via clathrin-mediated endocytosis...
September 22, 2017: Nature Communications
Gang Lu, Jia Li, Yijun Zhou, Xueping Zhou, Xiaorong Tao
Rice stripe tenuivirus (RSV) is a filamentous, negative-strand RNA virus causing severe diseases on rice in Asian countries. The viral particle is composed predominantly of a nucleocapsid protein (NP) and genomic RNA. However, the molecular details of how the RSV NP interacts with genomic RNA during particle assembly remain largely unknown. Here, we modeled the NP-RNA complex and show that polar amino acids within a predicted groove of NP are critical for RNA binding and protecting the RNA from RNase digestion...
June 2017: Virology
Alexander Ludwig, Tra Huong Nguyen, Daniel Leong, Laxmi Iyer Ravi, Boon Huan Tan, Sara Sandin, Richard J Sugrue
Respiratory syncytial virus (RSV) is an enveloped virus that assembles into filamentous virus particles on the surface of infected cells. Morphogenesis of RSV is dependent upon cholesterol-rich (lipid raft) membrane microdomains, but the specific role of individual raft molecules in RSV assembly is not well defined. Here, we show that RSV morphogenesis occurs within caveolar membranes and that both caveolin-1 and cavin-1 (also known as PTRF), the two major structural and functional components of caveolae, are actively recruited to and incorporated into the RSV envelope...
March 15, 2017: Journal of Cell Science
Shadi Shahriari, James Gordon, Reena Ghildyal
Respiratory syncytial virus (RSV) is one of the major pathogens responsible for lower respiratory tract infections (LRTI) in young children, the elderly, and the immunosuppressed. Currently, there are no antiviral drugs or vaccines available that effectively target RSV infections, proving a significant challenge in regards to prevention and treatment. An in-depth understanding of the host-virus interactions that underlie assembly and budding would inform new targets for antiviral development.Current research suggests that the polymerised form of actin, the filamentous or F-actin, plays a role in RSV assembly and budding...
September 26, 2016: Virology Journal
Tra Nguyen Huong, Laxmi Iyer Ravi, Boon Huan Tan, Richard J Sugrue
BACKGROUND: During respiratory syncytial virus (RSV) infection filamentous virus particles are formed on the cell surface. Although the virus infectivity remains cell-associated, low levels of cell-free virus is detected during advanced infection. It is currently unclear if this cell-free virus infectivity is due to a low-efficiency specific cell-release mechanism, or if it arises due to mechanical breakage following virus-induced cell damage at the advanced stage of infection. Understanding the origin of this cell-free virus is a prerequisite for understanding the mechanism of RSV transmission in permissive cells...
January 20, 2016: Virology Journal
Andreas Förster, Goedele N Maertens, Paul J Farrell, Monika Bajorek
UNLABELLED: Respiratory syncytial virus (RSV) infects epithelial cells of the respiratory tract and is a major cause of bronchiolitis and pneumonia in children and the elderly. The virus assembles and buds through the plasma membrane, forming elongated membrane filaments, but details of how this happens remain obscure. Oligomerization of the matrix protein (M) is a key step in the process of assembly and infectious virus production. In addition, it was suggested to affect the conformation of the fusion protein, the major current target for RSV antivirals, in the mature virus...
April 2015: Journal of Virology
Muhammad Raihan Jumat, Tra Ngyen Huong, Laxmi Iyer Ravi, Rebecca Stanford, Boon Huan Tan, Richard J Sugrue
We examined the effect of respiratory syncytial virus (RSV) infection on viperin protein expression in the permissive HEp2 and non-permissive RAW 264.7 macrophage cell lines. In RSV-infected HEp2 cells low levels of the viperin protein was localized to the virus-induced inclusion bodies and did not impair virus transmission in these cells. In contrast, RSV-infected RAW 264.7 cells increased expression of the STAT1 protein occurred at between 6 and 12h post-infection, which coincided with the appearance of P-STAT1...
February 2015: Antiviral Research
Gabriella Kiss, Jens M Holl, Grant M Williams, Eric Alonas, Daryll Vanover, Aaron W Lifland, Manasa Gudheti, Ricardo C Guerrero-Ferreira, Vinod Nair, Hong Yi, Barney S Graham, Philip J Santangelo, Elizabeth R Wright
UNLABELLED: Respiratory syncytial virus (RSV), a member of the Paramyxoviridae family of nonsegmented, negative-sense, single-stranded RNA genome viruses, is a leading cause of lower respiratory tract infections in infants, young children, and the elderly or immunocompromised. There are many open questions regarding the processes that regulate human RSV (hRSV) assembly and budding. Here, using cryo-electron tomography, we identified virus particles that were spherical, filamentous, and asymmetric in structure, all within the same virus preparation...
July 2014: Journal of Virology
M Bajorek, L Caly, K C Tran, G N Maertens, R A Tripp, E Bacharach, M N Teng, R Ghildyal, D A Jans
UNLABELLED: Human respiratory syncytial virus (RSV) is the most common cause of bronchiolitis and pneumonia in infants and the elderly worldwide; however, there is no licensed RSV vaccine or effective drug treatment available. The RSV matrix (M) protein plays key roles in virus assembly and budding, but the protein interactions that govern budding of infectious virus are not known. In this study, we focus on M protein and identify a key phosphorylation site (Thr205) in M that is critical for RSV infectious virus production...
June 2014: Journal of Virology
Yan Huo, Wenwen Liu, Fujie Zhang, Xiaoying Chen, Li Li, Qifei Liu, Yijun Zhou, Taiyun Wei, Rongxiang Fang, Xifeng Wang
Most plant viruses are transmitted by hemipteroid insects. Some viruses can be transmitted from female parent to offspring usually through eggs, but the mechanism of this transovarial transmission remains unclear. Rice stripe virus (RSV), a Tenuivirus, transmitted mainly by the small brown planthopper (Laodelphax striatellus), is also spread to the offspring through the eggs. Here, we used the RSV-planthopper system as a model to investigate the mechanism of transovarial transmission and demonstrated the central role of vitellogenin (Vg) of L...
March 2014: PLoS Pathogens
Min Yao, Xiaofan Liu, Shuo Li, Yi Xu, Yijun Zhou, Xueping Zhou, Xiaorong Tao
UNLABELLED: The NSvc2 glycoproteins encoded by Rice stripe tenuivirus (RSV) share many characteristics common to the glycoproteins found among Bunyaviridae. Within this viral family, glycoproteins targeting to the Golgi apparatus play a pivotal role in the maturation of the enveloped spherical particles. RSV particles, however, adopt a long filamentous morphology. Recently, RSV NSvc2 glycoproteins were shown to localize exclusively to the ER in Sf9 insect cells. Here, we demonstrate that the amino-terminal NSvc2 (NSvc2-N) targets to the Golgi apparatus in Nicotiana benthamiana cells, whereas the carboxyl-terminal NSvc2 (NSvc2-C) accumulates in the endoplasmic reticulum (ER)...
March 2014: Journal of Virology
Laxmi Iyer Ravi, Li Liang, Pui San Wong, Gaie Brown, Boon Huan Tan, Richard J Sugrue
We have examined the role that hydroxymethylglutaryl coenzyme A reductase (HMGCR) plays during respiratory syncytial virus (RSV) maturation. Imaging analysis indicated that virus-induced changes in F-actin structure correlated with the formation of virus filaments, and that these virus filaments played a direct role in virus cell-to-cell transmission. Treatment with cytochalasin D (CYD) prevented virus filament formation and virus transmission, but this could be reversed by removal of CYD. This observation, together with the presence of F-actin within the virus filaments suggested that newly polymerised F-actin was required for virus transmission...
October 2013: Antiviral Research
Yuanyuan Ma, Wei Wu, Hongyan Chen, Qifei Liu, Dongsheng Jia, Qianzhuo Mao, Qian Chen, Zujian Wu, Taiyun Wei
A cell line from the small brown planthopper (SBPH; Laodelphax striatellus) was established to study replication of rice stripe virus (RSV), a tenuivirus. The SBPH cell line, which had been subcultured through 30 passages, formed monolayers of epithelial-like cells. Inoculation of cultured vector cells with RSV resulted in a persistent infection. During viral infection in the SBPH cell line, the viral non-structural protein NS3 co-localized with the filamentous ribonucleoprotein particles of RSV, as revealed by electron and confocal microscopy...
June 2013: Journal of General Virology
Jun Fuchimoto, Takashi Kojima, Tamaki Okabayashi, Tomoyuki Masaki, Noriko Ogasawara, Kazufumi Obata, Kazuaki Nomura, Satoshi Hirakawa, Naoyuki Kobayashi, Tatsuro Shigyo, Shin-ichi Yokota, Nobuhiro Fujii, Hiroyuki Tsutsumi, Tetsuo Himi, Norimasa Sawada
Respiratory syncytial virus (RSV) is the major infectious agent causing serious respiratory tract inflammation in infants and young children. However, an effective vaccine and anti-viral therapy for RSV infection have not yet been developed. Hop-derived bitter acids have potent pharmacological effects on inflammation. Therefore, we investigated the effects of humulone, which is the main constituent of hop bitter acids, on the replication of RSV and release of the proinflammatory cytokine IL-8 and chemokine RANTES in RSV-infected human nasal epithelial cells (HNECs)...
December 2013: Medical Molecular Morphology
Yi Xu, Xueping Zhou
Our previous work has demonstrated that the NSvc4 protein of Rice stripe virus (RSV) functions as a cell-to-cell movement protein. However, the mechanisms whereby RSV traffics through plasmodesmata (PD) are unknown. Here we provide evidence that the NSvc4 moves on the actin filament and endoplasmic reticulum network, but not microtubules, to reach cell wall PD. Disruption of cytoskeleton using different inhibitors altered NSvc4 localization to PD, thus impeding RSV infection of Nicotiana benthamiana. Sequence analyses and deletion mutagenesis experiment revealed that the N-terminal 125 amino acids (AAs) of the NSvc4 determine PD targeting and that a transmembrane domain spanning AAs 106-125 is critical for PD localization...
2012: Frontiers in Plant Science
Fyza Y Shaikh, Thomas J Utley, Ryan E Craven, Meredith C Rogers, Lynne A Lapierre, James R Goldenring, James E Crowe
Respiratory syncytial virus (RSV) is a single-stranded RNA virus that assembles into viral filaments at the cell surface. Virus assembly often depends on the ability of a virus to use host proteins to accomplish viral tasks. Since the fusion protein cytoplasmic tail (FCT) is critical for viral filamentous assembly, we hypothesized that host proteins important for viral assembly may be recruited by the FCT. Using a yeast two-hybrid screen, we found that filamin A interacted with FCT, and mammalian cell experiments showed it localized to viral filaments but did not affect viral replication...
2012: PloS One
Fyza Y Shaikh, Reagan G Cox, Aaron W Lifland, Anne L Hotard, John V Williams, Martin L Moore, Philip J Santangelo, James E Crowe
UNLABELLED: Respiratory syncytial virus (RSV) is a single-stranded RNA virus in the Paramyxoviridae family that assembles into filamentous structures at the apical surface of polarized epithelial cells. These filaments contain viral genomic RNA and structural proteins, including the fusion (F) protein, matrix (M) protein, nucleoprotein (N), and phosphoprotein (P), while excluding F-actin. It is known that the F protein cytoplasmic tail (FCT) is necessary for filament formation, but the mechanism by which the FCT mediates assembly into filaments is not clear...
2012: MBio
Rajni Kumaria, Laxmi Ravi Iyer, Martin L Hibberd, Eric A F Simões, Richard J Sugrue
BACKGROUND: Human respiratory syncytial virus (HRSV) is the most important virus causing lower respiratory infection in young children. The complete genetic characterization of RSV clinical strains is a prerequisite for understanding HRSV infection in the clinical context. Current information about the genetic structure of the HRSV genome has largely been obtained using tissue culture adapted viruses. During tissue culture adaptation genetic changes can be introduced into the virus genome, which may obscure subtle variations in the genetic structure of different RSV strains...
2011: Virology Journal
Mohammad Jamaluddin, John E Wiktorowicz, Kizhake V Soman, Istvan Boldogh, Jeffrey D Forbus, Heidi Spratt, Roberto P Garofalo, Allan R Brasier
The respiratory epithelium plays a central role in innate immunity by secreting networks of inflammatory mediators in response to respiratory syncytial virus (RSV) infection. Previous proteomic studies focusing on the host cellular response to RSV indicated the existence of a nuclear heat shock response and cytoplasmic depletion of antioxidant proteins in model type II-like airway epithelial cells. Here, we increased the depth of nuclear proteomic interrogation by using fluorescence difference labeling followed by liquid isoelectric focusing prefractionation/two-dimensional gel electrophoresis (2-DE) to identify an additional 41 proteins affected by RSV infection...
September 2010: Journal of Virology
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