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Sagar M. Goyal - One of the best experts on this subject based on the ideXlab platform.
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Forward and reverse primers with size and annealing temperatures.
2018Co-Authors: Hamada A. Aboubakr, Sunil K. Mor, Leeann Higgins, Anibal Armien, Mohammed M. Youssef, Peter J. Bruggeman, Sagar M. GoyalAbstract:The position of gel-based RT-PCR primers are based on USDA Feline Calicivirus sequence (GenBank accession AY560118.1).
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Genomic characterization of a novel Calicivirus, FHMCV-2012, from baitfish in the USA
Archives of virology, 2017Co-Authors: Sunil K. Mor, Nicholas B. D. Phelps, Kuttichantran Subramaniam, Alexander E Primus, Aníbal G. Armién, Rebekah Mccann, Corey Puzach, Thomas B. Waltzek, Sagar M. GoyalAbstract:During regulatory sampling of fathead minnows (Pimephales promelas), a novel Calicivirus was isolated from homogenates of kidney and spleen inoculated into bluegill fry (BF-2) cells. Infected cell cultures exhibiting cytopathic effects were screened by PCR-based methods for selected fish viral pathogens. Illumina HiSeq next generation sequencing of the total RNA revealed a novel Calicivirus genome that showed limited protein sequence similarity to known homologs in a BLASTp search. The complete genome of this fathead minnow Calicivirus (FHMCV) is 6564 nt long, encoding a polyprotein of 2114 aa in length. The complete polyprotein shared only 21% identity with Atlantic salmon Calicivirus,followed by 11% to 14% identity with mammalian Caliciviruses. A molecular detection assay (RT-PCR) was designed from this sequence for screening of field samples for FHMCV in the future. This virus likely represents a prototype species of a novel genus in the family Caliciviridae, tentatively named “Minovirus”.
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Effect of temperature and sanitizers on the survival of feline Calicivirus, Escherichia coli, and F-specific coliphage MS2 on leafy salad vegetables.
Journal of food protection, 2004Co-Authors: Paul B. Allwood, Yashpal Singh Malik, Craig W. Hedberg, Sagar M. GoyalAbstract:We conducted a series of experiments to compare the survival of Escherichia coli, feline Calicivirus, and F-specific coliphage MS2 on lettuce and cabbage with and without disinfection. Inoculated produce was held at 4, 25, or 37°C for 21 days or was treated with different concentrations of sodium bicarbonate, chlorine bleach, peroxyacetic acid, or hydrogen peroxide. Survival was measured by the decimal reduction value (time to 90% reduction in titer) and the change in log titers of the test organisms. A stronger correlation of survival measures was observed between feline Calicivirus and MS2 than between E. coli and either of the viral agents at 25 and 37°C. The maximum time to detection limit for MS2 at all temperatures was 9 days, whereas feline Calicivirus was detected for a maximum of 14 days at 4°C. In contrast, E. coli was detectable for 21 days at 4 and 25°C and for 14 days at 37°C. Significant increases in E. coli titer occurred within the first 5 days, but virus titers decreased steadily througho...
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effect of temperature and sanitizers on the survival of feline Calicivirus escherichia coli and f specific coliphage ms2 on leafy salad vegetables
Journal of Food Protection, 2004Co-Authors: Paul B. Allwood, Yashpal Singh Malik, Craig W. Hedberg, Sagar M. GoyalAbstract:We conducted a series of experiments to compare the survival of Escherichia coli, feline Calicivirus, and F-specific coliphage MS2 on lettuce and cabbage with and without disinfection. Inoculated produce was held at 4, 25, or 37 degrees C for 21 days or was treated with different concentrations of sodium bicarbonate, chlorine bleach, peroxyacetic acid, or hydrogen peroxide. Survival was measured by the decimal reduction value (time to 90% reduction in titer) and the change in log titers of the test organisms. A stronger correlation of survival measures was observed between feline Calicivirus and MS2 than between E. coli and either of the viral agents at 25 and 37 degrees C. The maximum time to detection limit for MS2 at all temperatures was 9 days, whereas feline Calicivirus was detected for a maximum of 14 days at 4 degrees C. In contrast, E. coli was detectable for 21 days at 4 and 25 degrees C and for 14 days at 37 degrees C. Significant increases in E. coli titer occurred within the first 5 days, but virus titers decreased steadily throughout the experiments. E. coli was also highly susceptible to all disinfectants except 1% sodium bicarbonate and 50 ppm chlorine bleach, whereas the viruses were resistant to all four disinfectants.
Ian Goodfellow - One of the best experts on this subject based on the ideXlab platform.
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protein rna linkage and posttranslational modifications of feline Calicivirus and murine norovirus vpg proteins
PeerJ, 2016Co-Authors: Allan Olspert, Yasmin Chaudhry, Myra Hosmillo, Lauri Peil, Erkki Truve, Ian GoodfellowAbstract:Members of the Caliciviridae family of positive sense RNA viruses cause a wide range of diseases in both humans and animals. The detailed characterization of the Calicivirus life cycle had been hampered due to the lack of robust cell culture systems and experimental tools for many of the members of the family. However, a number of Caliciviruses replicate efficiently in cell culture and have robust reverse genetics systems available, most notably feline Calicivirus (FCV) and murine norovirus (MNV). These are therefore widely used as representative members with which to examine the mechanistic details of Calicivirus genome translation and replication. The replication of the Calicivirus RNA genome occurs via a double-stranded RNA intermediate that is then used as a template for the production of new positive sense viral RNA, which is covalently linked to the virus-encoded protein VPg. The covalent linkage to VPg occurs during genome replication via the nucleotidylylation activity of the viral RNA-dependent RNA polymerase. Using FCV and MNV, we used mass spectrometry-based approach to identify the specific amino acid linked to the 5' end of the viral nucleic acid. We observed that both VPg proteins are covalently linked to guanosine diphosphate (GDP) moieties via tyrosine positions 24 and 26 for FCV and MNV respectively. These data fit with previous observations indicating that mutations introduced into these specific amino acids are deleterious for viral replication and fail to produce infectious virus. In addition, we also detected serine phosphorylation sites within the FCV VPg protein with positions 80 and 107 found consistently phosphorylated on VPg-linked viral RNA isolated from infected cells. This work provides the first direct experimental characterization of the linkage of infectious Calicivirus viral RNA to the VPg protein and highlights that post-translational modifications of VPg may also occur during the viral life cycle.
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Functions of the 5' and 3' ends of Calicivirus genomes
Virus research, 2015Co-Authors: Bader Alhatlani, Surender Vashist, Ian GoodfellowAbstract:The Caliciviridae family of small positive sense RNA viruses contains a diverse range of pathogens of both man and animals. The molecular mechanisms of Calicivirus genome replication and translation have not been as widely studied as many other RNA viruses. With the relatively recent development of robust cell culture and reverse genetics systems for several members of the Caliciviridae family, a more in-depth analysis of the finer detail of the viral life cycle has now been obtained. As a result, the identification and characterization of the role of RNA structures in the Calicivirus life cycle has also been possible. This review aims to summarize the current state of knowledge with respect to the role of RNA structures at the termini of Calicivirus genomes.
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nucleolin interacts with the feline Calicivirus 3 untranslated region and the protease polymerase ns6 and ns7 proteins playing a role in virus replication
Journal of Virology, 2011Co-Authors: Clotilde Canciolonches, Luis Urena, Surender Vashist, Ian Goodfellow, Martha Yocupiciomonroy, Carlos Sandovaljaime, Ivan Galvanmendoza, Juan Santiago Salasbenito, Ana Lorena GutierrezescolanoAbstract:Cellular proteins play many important roles during the life cycle of all viruses. Specifically, host cell nucleic acid-binding proteins interact with viral components of positive-stranded RNA viruses and regulate viral translation, as well as RNA replication. Here, we report that nucleolin, a ubiquitous multifunctional nucleolar shuttling phosphoprotein, interacts with the Norwalk virus and feline Calicivirus (FCV) genomic 3 untranslated regions (UTRs). Nucleolin can also form a complex in vitro with recombinant Norwalk virus NS6 and -7 (NS6/7) and can be copurified with the analogous protein from feline Calicivirus (p76 or NS6/7) from infected feline kidney cells. Nucleolin RNA levels or protein were not modified during FCV infection; however, as a consequence of the infection, nucleolin was seen to relocalize from the nucleoli to the nucleoplasm, as well as to the perinuclear area where it colocalizes with the feline Calicivirus NS6/7 protein. In addition, antibodies to nucleolin were able to precipitate viral RNA from feline Calicivirus-infected cells, indicating a direct or indirect association of nucleolin with the viral RNA during virus replication. Small interfering RNA (siRNA)-mediated knockdown of nucleolin resulted in a reduction of the cytopathic effect and virus yield in CrFK cells. Taken together, these results demonstrate that nucleolin is a nucleolar component that interacts with viral RNA and NS6/7 and is required for feline Calicivirus replication.
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Nucleolin Interacts with the Feline Calicivirus 3′ Untranslated Region and the Protease-Polymerase NS6 and NS7 Proteins, Playing a Role in Virus Replication
Journal of virology, 2011Co-Authors: Clotilde Cancio-lonches, Martha Yocupicio-monroy, Carlos Sandoval-jaime, Iván Galván-mendoza, Luis Urena, Surender Vashist, Ian Goodfellow, Juan Santiago Salas-benito, Ana Lorena Gutiérrez-escolanoAbstract:Cellular proteins play many important roles during the life cycle of all viruses. Specifically, host cell nucleic acid-binding proteins interact with viral components of positive-stranded RNA viruses and regulate viral translation, as well as RNA replication. Here, we report that nucleolin, a ubiquitous multifunctional nucleolar shuttling phosphoprotein, interacts with the Norwalk virus and feline Calicivirus (FCV) genomic 3' untranslated regions (UTRs). Nucleolin can also form a complex in vitro with recombinant Norwalk virus NS6 and -7 (NS6/7) and can be copurified with the analogous protein from feline Calicivirus (p76 or NS6/7) from infected feline kidney cells. Nucleolin RNA levels or protein were not modified during FCV infection; however, as a consequence of the infection, nucleolin was seen to relocalize from the nucleoli to the nucleoplasm, as well as to the perinuclear area where it colocalizes with the feline Calicivirus NS6/7 protein. In addition, antibodies to nucleolin were able to precipitate viral RNA from feline Calicivirus-infected cells, indicating a direct or indirect association of nucleolin with the viral RNA during virus replication. Small interfering RNA (siRNA)-mediated knockdown of nucleolin resulted in a reduction of the cytopathic effect and virus yield in CrFK cells. Taken together, these results demonstrate that nucleolin is a nucleolar component that interacts with viral RNA and NS6/7 and is required for feline Calicivirus replication.
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Calicivirus translation initiation requires an interaction between VPg and eIF4E
EMBO reports, 2005Co-Authors: Ian Goodfellow, Yasmin Chaudhry, Ioanna Gioldasi, Andreas Gerondopoulos, Alessandro Natoni, Louisette Labrie, Jean-françois Laliberté, Lisa O. RobertsAbstract:Unlike other positive-stranded RNA viruses that use either a 5'-cap structure or an internal ribosome entry site to direct translation of their messenger RNA, Calicivirus translation is dependent on the presence of a protein covalently linked to the 5' end of the viral genome (VPg). We have shown a direct interaction of the Calicivirus VPg with the cap-binding protein eIF 4 E. This interaction is required for Calicivirus mRNA translation, as sequestration of eIF 4 E by 4 E-BP 1 inhibits translation. Functional analysis has shown that VPg does not interfere with the interaction between eIF 4 E and the cap structure or 4 E-BP 1, suggesting that VPg binds to eIF 4 E at a different site from both cap and 4 E-BP 1. This work lends support to the idea that Calicivirus VPg acts as a novel 'cap substitute' during initiation of translation on virus mRNA.
Kim Y. Green - One of the best experts on this subject based on the ideXlab platform.
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visualization of feline Calicivirus replication in real time with recombinant viruses engineered to express fluorescent reporter proteins
Virology, 2010Co-Authors: Eugenio J Abente, Stanislav V. Sosnovtsev, Kim Y. Green, Karin BokAbstract:Caliciviruses are non-enveloped, icosahedral viruses with a single-stranded, positive sense RNA genome. Transposon-mediated insertional mutagenesis was used to insert a transprimer sequence into random sites of an infectious full-length cDNA clone of the feline Calicivirus (FCV) genome. A site in the LC gene (encoding the capsid leader protein) of the FCV genome was identified that could tolerate foreign insertions, and two viable recombinant FCV variants expressing LC fused either to AcGFP, or DsRedFP were recovered. The effects of the insertions on LC processing, RNA replication, and stability of the viral genome were analyzed, and the progression of a Calicivirus single infection and co-infection were captured by real-time imaging fluorescent microscopy. The ability to engineer viable recombinant Caliciviruses expressing foreign markers enables new approaches to investigate virus and host cell interactions, as well as studies of viral recombination, one of the driving forces of Calicivirus evolution.
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Calicivirus 3C-Like Proteinase Inhibits Cellular Translation by Cleavage of Poly(A)-Binding Protein
Journal of Virology, 2004Co-Authors: Muge N. Kuyumcu-martinez, Stanislav V. Sosnovtsev, Kim Y. Green, Gaël Belliot, Kyeong-ok Chang, Richard E. LloydAbstract:Caliciviruses are single-stranded RNA viruses that cause a wide range of diseases in both humans and animals, but little is known about the regulation of cellular translation during infection. We used two distinct Calicivirus strains, MD145-12 (genus Norovirus) and feline Calicivirus (FCV) (genus Vesivirus), to investigate potential strategies used by the Caliciviruses to inhibit cellular translation. Recombinant 3C-like proteinases (r3CL pro ) from norovirus and FCV were found to cleave poly(A)-binding protein (PABP) in the absence of other viral proteins. The norovirus r3CL pro PABP cleavage products were indistinguishable from those generated by poliovirus (PV) 3C pro cleavage, while the FCV r3CL pro products differed due to cleavage at an alternate cleavage site 24 amino acids downstream of one of the PV 3C pro cleavage sites. All cleavages by Calicivirus or PV proteases separated the C-terminal domain of PABP that binds translation factors eIF4B and eRF3 from the N-terminal RNA-binding domain of PABP. The effect of PABP cleavage by the norovirus r3CL pro was analyzed in HeLa cell translation extracts, and the presence of r3CL pro inhibited translation of both endogenous and exogenous mRNAs. Translation inhibition was poly(A) dependent, and replenishment of the extracts with PABP restored translation. Analysis of FCV-infected feline kidney cells showed that the levels of de novo cellular protein synthesis decreased over time as virus-specific proteins accumulated, and cleavage of PABP occurred in virus-infected cells. Our data indicate that the Calicivirus 3CL pro , like PV 3C pro , mediates the cleavage of PABP as part of its strategy to inhibit cellular translation. PABP cleavage may be a common mechanism among certain virus families to manipulate cellular translation. The family Caliciviridae is comprised of four genera: Vesivirus (including feline Calicivirus [FCV]), Lagovirus (including rabbit hemorrhagic disease virus [RHDV]), Norovirus (including Norwalk virus and MD145-12), and Sapovirus (including Sapporo virus and porcine enteric Calicivirus [PEC]) (21). They are single-stranded, positive-sense RNA viruses that share evolutionary relatedness with the picornaviruses. The major Caliciviruses associated with human disease belong to the genus Norovirus. The noroviruses (designated here as NoV) are responsible for the majority of food- and waterborne outbreaks of viral gastroenteritis worldwide (15, 26), but the development of control strategies for NoV disease has been complicated by the absence of tissue culture systems to study replication (23). Analysis of Calicivirus proteins in comparative studies with the picornaviruses might yield insight into their functions during viral replication and facilitate the development of treatment strategies for Calicivirus-associated diseases. In this study, we investigated possible strategies that calici
Sabah Bidawid - One of the best experts on this subject based on the ideXlab platform.
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Survival of Calicivirus in foods and on surfaces: experiments with feline Calicivirus as a surrogate for norovirus.
Journal of Food Protection, 2007Co-Authors: Kirsten Mattison, Michael Abebe, J.m. Farber, K Karthikeyan, N Malik, Syed A Sattar, Sabah BidawidAbstract:Although there is a large body of evidence incriminating foods as vehicles in the transmission of norovirus, little is known about virus survival in foods and on surfaces. Feline Calicivirus was us...
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Survival of Calicivirus in foods and on surfaces: experiments with feline Calicivirus as a surrogate for norovirus.
Journal of food protection, 2007Co-Authors: Kirsten Mattison, Michael Abebe, J.m. Farber, K Karthikeyan, N Malik, Syed A Sattar, Sabah BidawidAbstract:Although there is a large body of evidence incriminating foods as vehicles in the transmission of norovirus, little is known about virus survival in foods and on surfaces. Feline Calicivirus was used as a surrogate for norovirus to investigate its survival in representative foods of plant and animal origin and on metal surfaces. Known concentrations of feline Calicivirus in a natural fecal suspension were deposited onto lettuce, strawberries, ham, or stainless steel and incubated for 7 days at refrigeration or room temperatures. Virus was recovered at 1-day intervals, and the titers of the virus were determined by plaque assay. Infectious virus was recoverable until day 7 from lettuce, ham, and stainless steel. Statistically higher titers of feline Calicivirus (P < 0.05) were recovered from ham under all conditions than from lettuce, strawberries, or stainless steel. These data provide valuable information for epidemiological and monitoring purposes as well as for the development of food processing practices and appropriate strategies to inactivate norovirus and control its transmission via foods and surfaces.
Paul B. Allwood - One of the best experts on this subject based on the ideXlab platform.
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Effect of temperature and sanitizers on the survival of feline Calicivirus, Escherichia coli, and F-specific coliphage MS2 on leafy salad vegetables.
Journal of food protection, 2004Co-Authors: Paul B. Allwood, Yashpal Singh Malik, Craig W. Hedberg, Sagar M. GoyalAbstract:We conducted a series of experiments to compare the survival of Escherichia coli, feline Calicivirus, and F-specific coliphage MS2 on lettuce and cabbage with and without disinfection. Inoculated produce was held at 4, 25, or 37°C for 21 days or was treated with different concentrations of sodium bicarbonate, chlorine bleach, peroxyacetic acid, or hydrogen peroxide. Survival was measured by the decimal reduction value (time to 90% reduction in titer) and the change in log titers of the test organisms. A stronger correlation of survival measures was observed between feline Calicivirus and MS2 than between E. coli and either of the viral agents at 25 and 37°C. The maximum time to detection limit for MS2 at all temperatures was 9 days, whereas feline Calicivirus was detected for a maximum of 14 days at 4°C. In contrast, E. coli was detectable for 21 days at 4 and 25°C and for 14 days at 37°C. Significant increases in E. coli titer occurred within the first 5 days, but virus titers decreased steadily througho...
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effect of temperature and sanitizers on the survival of feline Calicivirus escherichia coli and f specific coliphage ms2 on leafy salad vegetables
Journal of Food Protection, 2004Co-Authors: Paul B. Allwood, Yashpal Singh Malik, Craig W. Hedberg, Sagar M. GoyalAbstract:We conducted a series of experiments to compare the survival of Escherichia coli, feline Calicivirus, and F-specific coliphage MS2 on lettuce and cabbage with and without disinfection. Inoculated produce was held at 4, 25, or 37 degrees C for 21 days or was treated with different concentrations of sodium bicarbonate, chlorine bleach, peroxyacetic acid, or hydrogen peroxide. Survival was measured by the decimal reduction value (time to 90% reduction in titer) and the change in log titers of the test organisms. A stronger correlation of survival measures was observed between feline Calicivirus and MS2 than between E. coli and either of the viral agents at 25 and 37 degrees C. The maximum time to detection limit for MS2 at all temperatures was 9 days, whereas feline Calicivirus was detected for a maximum of 14 days at 4 degrees C. In contrast, E. coli was detectable for 21 days at 4 and 25 degrees C and for 14 days at 37 degrees C. Significant increases in E. coli titer occurred within the first 5 days, but virus titers decreased steadily throughout the experiments. E. coli was also highly susceptible to all disinfectants except 1% sodium bicarbonate and 50 ppm chlorine bleach, whereas the viruses were resistant to all four disinfectants.
