Mouse Norovirus 1 Utilizes the Cytoskeleton Network To Establish Localization of the Replication Complex Proximal to the Microtubule Organizing Center

Hyde, Jennifer; Gillespie, Leah; Mackenzie, Jason M.

doi:10.1128/jvi.05784-11

Cited by 48 publications

(54 citation statements)

References 56 publications

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“…As previously reported (30), NS4-FLAG colocalized with NS7 in dense perinuclear foci (Fig. 8), in a position most likely consistent with the microtubule-organizing center, which has recently been reported to be the location for the replication complex (29). NS7 also appeared to show some diffuse staining throughout the cytoplasm of all infected cells, as has previously been observed (30).…”

Section: Figsupporting

confidence: 63%

High-Resolution Functional Profiling of the Norovirus Genome

Thorne

Bailey

Goodfellow

2012

J Virol

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Human noroviruses (HuNoV) are a major cause of nonbacterial gastroenteritis worldwide, yet details of the life cycle and replication of HuNoV are relatively unknown due to the lack of an efficient cell culture system. Studies with murine norovirus (MNV), which can be propagated in permissive cells, have begun to probe different aspects of the norovirus life cycle; however, our understanding of the specific functions of the viral proteins lags far behind that of other RNA viruses. Genome-wide functional profiling by insertional mutagenesis can reveal protein domains essential for replication and can lead to generation of tagged viruses, which has not yet been achieved for noroviruses. Here, transposon-mediated insertional mutagenesis was used to create 5 libraries of mutagenized MNV infectious clones, each containing a 15-nucleotide sequence randomly inserted within a defined region of the genome. Infectious virus was recovered from each library and was subsequently passaged in cell culture to determine the effect of each insertion by insertion-specific fluorescent PCR profiling. Genome-wide profiling of over 2,000 insertions revealed essential protein domains and confirmed known functional motifs. As validation, several insertion sites were introduced into a wild-type clone, successfully allowing the recovery of infectious virus. Screening of a number of reporter proteins and epitope tags led to the generation of the first infectious epitope-tagged noroviruses carrying the FLAG epitope tag in either NS4 or VP2. Subsequent work confirmed that epitope-tagged fully infectious noroviruses may be of use in the dissection of the molecular interactions that occur within the viral replication complex. Human norovirus (HuNoV) is the leading cause of nonbacterial gastroenteritis in developed countries, with initial reports also indicating high incidence and mortality in developing countries (47). Due to the low infectious dose, multiple transmission routes, and high stability of HuNoV in the environment (41, 72), outbreaks typically occur in places with close living conditions, such as hospitals. Infections in hospitals alone cost the United Kingdom National Health Service an estimated £115 million and result in 47,000 lost bed days per year, compromising hospital services (39, 53). The majority of HuNoV infections are acute and self-resolving; however, links with conditions such as inflammatory bowel disease (36), infantile seizures (18), and neonatal necrotizing enterocolitis (65, 74) have been established. There are also increasing reports of persistent infections in the elderly and the immunocompromised, such as transplant recipients and some cancer patients (2,13,20,40,55,56).Details of the life cycle and replication of HuNoV are relatively unknown due to the lack of an efficient cell culture system. The discovery of murine norovirus (MNV) in 2003 has since facilitated studies on norovirus replication. Unlike HuNoV, MNV can be propagated in culture, displaying a tropism for macrophage and dendritic cells (75), and ca...

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Section: Figsupporting

confidence: 63%

High-Resolution Functional Profiling of the Norovirus Genome

Thorne

Bailey

Goodfellow

2012

J Virol

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“…To date, a direct nucleic acidbinding activity of Talin-1 has not been reported; however, this protein has been found associated with a number of nucleic acidbinding proteins also identified in this study, namely, YBX1, FUS, hnRNPK, hnRNP E1 (PCBP1), and PABP (13). Recent studies indicated that MNV replication requires components of the cytoskeleton network for correct localization of the viral replication complex (29). Further studies to dissect a possible role for Talin-1 in the norovirus life cycle are under way.…”

Section: Figmentioning

confidence: 91%

Identification of RNA-Protein Interaction Networks Involved in the Norovirus Life Cycle

Vashist

Ureña

Chaudhry

et al. 2012

J Virol

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Human noroviruses are one of the major causes of acute gastroenteritis in the developed world, yet our understanding of their molecular mechanisms of genome translation and replication lags behind that for many RNA viruses. Due to the nonculturable nature of human noroviruses, many related members of the Caliciviridae family of small RNA viruses are often used as model systems to dissect the finer details of the norovirus life cycle. Murine norovirus (MNV) has provided one such system with which to study the basic mechanisms of norovirus translation and replication in cell culture. In this report we describe the use of riboproteomics to identify host factors that interact with the extremities of the MNV genome. This network of RNA-protein interactions contains many well-characterized host factors, including PTB, La, and DDX3, which have been shown to play a role in the life cycle of other RNA viruses. By using RNA coimmunoprecipitation, we confirmed that a number of the factors identified using riboproteomics are associated with the viral RNA during virus replication in cell culture. We further demonstrated that RNA inhibition-mediated knockdown of the intracellular levels of a number of these factors inhibits or slows norovirus replication in cell culture, allowing identification of new intracellular targets for this important group of pathogens. The genomes of small positive-strand RNA viruses must not only encode the viral proteins required for viral genome replication and infectious virus production, but they must also contain cis-acting RNA sequences and structures that play critical roles in the virus life cycle (40). Host cell factors that interact with these RNA elements play important roles in all aspects of the virus life cycle (49); consequently, the manipulation of these interactions via the introduction of mutations into the viral genome is a viable approach to rational attenuation (24,26,57,78). In addition, the modification of these RNA-protein interactions by using small-molecule or peptide inhibitors has been shown to provide antiviral activity, at least in cell culture (8,19).Noroviruses, first identified in 1972 by Albert Kapikian (33), are now accepted as the major cause of viral gastroenteritis in the developed world (21). Reports indicate that in excess of 23 million cases occur each year in the United States, resulting in a significant economic impact. Now over 20 years since the first full genome sequence of a norovirus became available (77), investigators are still unable to efficiently propagate human noroviruses in cell culture (15, 37). Recent data have indicated that the viral RNA purified from the feces of individuals infected with Norwalk virus, the prototype human norovirus, can replicate when transfected into immortalized cells in culture (23). These data clearly demonstrate that one of the major blocks in the ability of human noroviruses to replicate in cell culture is the ability of the virus to enter and spread from cell to cell. It is also clear that the intracellular environ...

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“…Tubulin acetylation is such a modification that is involved in the replication process of several viruses, including herpes simplex virus 1 (HSV-1) [81], mouse norovirus 1 [82] and adenovirus [83]. Besides, tubulin acetylation is supposed to guide and accelerate the nuclear trafficking of post-entry viruses, and inhibition of HDAC6 by its specific inhibitors could apparently improve the replication efficiency of oncolytic viruses [84,85].…”

Section: Replicationmentioning

confidence: 99%

Cellular defence or viral assist: the dilemma of HDAC6

Zheng

Jiang

et al. 2017

Journal of General Virology

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Histone deacetylase 6 (HDAC6) is a unique cytoplasmic deacetylase that regulates various important biological processes by preventing protein aggregation and deacetylating different non-histone substrates including tubulin, heat shock protein 90, cortactin, retinoic acid inducible gene I and b-catenin. Growing evidence has indicated a dual role for HDAC6 in viral infection and pathogenesis: HDAC6 may represent a host defence mechanism against viral infection by modulating microtubule acetylation, triggering antiviral immune response and stimulating protective autophagy, or it may be hijacked by the virus to enhance proinflammatory response. In this review, we will highlight current data illustrating the complexity and importance of HDAC6 in viral pathogenesis. We will summarize the structure and functional specificity of HDAC6, and its deacetylaseand ubiquitin-dependent activity in key cellular events in response to virus infection. We will also discuss how HDAC6 exerts its direct or indirect histone modification ability in viral lytic-latency switch.

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Mouse Norovirus 1 Utilizes the Cytoskeleton Network To Establish Localization of the Replication Complex Proximal to the Microtubule Organizing Center

Cited by 48 publications

References 56 publications

High-Resolution Functional Profiling of the Norovirus Genome

High-Resolution Functional Profiling of the Norovirus Genome

Identification of RNA-Protein Interaction Networks Involved in the Norovirus Life Cycle

Cellular defence or viral assist: the dilemma of HDAC6

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