A Stable HeLa Cell Line That Inducibly Expresses Poliovirus 2A
            <sup>pro</sup>
            : Effects on Cellular and Viral Gene Expression

Barco, Ángel; Feduchi, Elena; Carrasco, Luís

doi:10.1128/jvi.74.5.2383-2392.2000

Cited by 34 publications

(42 citation statements)

References 71 publications

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“…On the one hand, this protease abrogates host protein synthesis, but it can also stimulate PV translation (20,2,9). The findings described in the present work certainly support these opposing effects, since there is a correlation between eIF4G hydrolysis and the inhibition of cellular protein synthesis, whereas translation driven by EMCV IRES element is stimulated (high EMC-2A dose) or at least is not affected (low EMC-2A dose) under these conditions.…”

Section: Discussionsupporting

confidence: 73%

“…Thus, hydrolysis of eIF4GI occurs after eIF4GII cleavage. This method is much more efficient, rapid, and reliable than other methods previously described, such as HeLa cell lines that can be induced to express PV 2A pro , direct penetration of hybrid proteins that contain this protease, or coupled infection with recombinant vaccinia virus T7 and transfection with pTM1-2A (20,17,21).…”

Section: Discussionmentioning

confidence: 99%

“…Plasmids-The plasmids pKs-Luc, pTM1-Luc, pTM1-2C, and pTM1-2A and the plasmids pTM1-2AG60R, pTM1-2AG121E, pTM1-2AD135N, and pTM1-2AV119M, which encodes a PV 2A pro mutant, have been described previously (20,21).…”

Section: Methodsmentioning

confidence: 99%

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Differential Cleavage of eIF4GI and eIF4GII in Mammalian Cells

Castelló

Álvarez

Carrasco

2006

Journal of Biological Chemistry

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Two isoforms of the translation initiation factor eIF4G, eIF4GI and eIF4GII, have been described in eukaryotic cells. The exact function of each isoform during the initiation of protein synthesis is still under investigation. We have developed an efficient and reliable method of expressing poliovirus 2Apro, which differentially proteolyzes eIF4GI and eIF4GII in a timeand dose-dependent manner. This system is based on the electroporation of an in vitro transcribed mRNA that contains the encephalomyocarditis virus internal ribosome entry site followed by the sequence of poliovirus 2Apro. In contrast to HeLa cells, expression of this protease in BHK-21 cells induces delayed hydrolysis kinetics of eIF4GI with respect to eIF4GII. Moreover, under these conditions the polyadenylate binding protein is not cleaved. Interestingly, translation of de novo synthesized luciferase mRNA is highly dependent on eIF4GI integrity, whereas ongoing translation is inhibited at the same time as eIF4GII cleavage. Moreover, reinitiation of a preexisting mRNA translation after polysome run-off is dependent on the integrity of eIF4GII. Notably, de novo translation of heat shock protein 70 mRNA depends little on eIF4GI integrity but is more susceptible to eIF4GII hydrolysis. Finally, translation of an mRNA containing encephalomyocarditis virus internal ribosome entry site when the two isoforms of eIF4G are differentially hydrolyzed has been examined.

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Section: Discussionsupporting

confidence: 73%

Section: Discussionmentioning

confidence: 99%

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Differential Cleavage of eIF4GI and eIF4GII in Mammalian Cells

Castelló

Álvarez

Carrasco

2006

Journal of Biological Chemistry

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“…Enterovirus 2A pro processes the viral polyprotein (62), and it also cleaves a variety of host proteins, including the translation proteins eIF4GI (36), eIF4GII (43), and poly(A)-binding protein (27). Proteinase 2A pro has been linked to the inhibition of host translation that occurs during enterovirus infection (1,63) and is also involved in processes as varied as viral RNA replication (42), viral translation (33), and viral RNA stability (30). Interferons (IFNs) are pleiotropic cytokines that are well known for their role in the cellular antiviral response (4,35,57).…”

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confidence: 99%

Proteinase 2A^proIs Essential for Enterovirus Replication in Type I Interferon-Treated Cells

Morrison

Racaniello

2009

J Virol

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The Picornaviridae family comprises a diverse group of small RNA viruses that cause a variety of human and animal diseases. Some of these viruses are known to induce cleavage of components of the innate immune system and to inhibit steps in the interferon pathway that lead to the production of type I interferon. There has been no study of the effect of picornaviral infection on the events that occur after interferons have been produced. To determine whether members of the Enterovirus genus can antagonize the antiviral activity of interferon-stimulated genes (ISGs), we pretreated cells with alpha interferon (IFN-␣) and then infected the cells with poliovirus type 1, 2, or 3; enterovirus type 70; or human rhinovirus type 16. We found that these viruses were able to replicate in IFN- ␣

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“…P2 and P3 are cleaved into seven mature nonstructural proteins; of these, 2A and 3C are proteases (16). 2A plays a prominent role in shut-off of host protein synthesis after viral infection (17), and 3C performs most of the cleavages of the viral polyprotein. Because of the known interactions of the proteases with cellular functions, we hypothesized that expression of the 3C protease is sufficient for cytokine up-regulation in airway epithelial cells.…”

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confidence: 99%

Rhinovirus 16 3C Protease Induces Interleukin-8 and Granulocyte-Macrophage Colony-Stimulating Factor Expression in Human Bronchial Epithelial Cells

et al. 2004

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Rhinovirus (RV), a member of the Picornaviridae family, accounts for many virus-induced asthma exacerbations. RV induces airway cell chemokine expression both in vivo and in vitro. Because of the known interactions of proteases with cellular functions, we hypothesized that RV 3C protease is sufficient for cytokine up-regulation. A cDNA encoding RV16 3C protease was constructed by PCR amplification and transfected into 16HBE14oϪ human bronchial epithelial cells. 3C protease induced expression of both IL-8 and GM-CSF, as well as transcription from both the IL-8 and GM-CSF promoters. 3C expression also induced activator protein 1 and NF-B transcriptional activation. Finally, mutation of IL-8 promoter AP-1 and NF-B promoter sequences significantly reduced 3C-induced responses. Together, these data suggest expression of RV16 3C protease is RVs are the most common upper respiratory pathogens, inducing the majority of common colds worldwide. Viral infections trigger asthma in 80 to 85% of asthma exacerbations in children and 44% in adults (1, 2) RV, a member of the Picornaviridae family of small, positive-stranded RNA viruses, accounts for much of virus-induced asthma exacerbations (1, 2). Cytokines are elaborated in vivo in the nasal mucosa, and most likely also from bronchial epithelium, after RV infection. Neutrophils, eosinophils, and lymphocytes increase in the nasal secretions of RV-infected patients compared with uninfected controls (3, 4).The cytokines IL-8 and GM-CSF have been implicated in the pathogenesis of asthma. IL-8 is a potent activator and chemoattractant of neutrophils (5) and eosinophils (6), and GM-CSF promotes the differentiation and survival of recruited eosinophils (7). Increased levels of IL-8, GM-CSF (8), and ICAM-1 (9), the receptor of 90% of RV serotypes, as well as IL-1 and IL-6, have been found in the nasal secretions of individuals infected with RVs (10 -12). In bronchial mucosal biopsies obtained after experimental infection with RV, Bardin and colleagues (13) noted a significant increase in submucosal lymphocytes and epithelial eosinophils, which, in asthmatics, persisted into convalescence. Although increases in cytokine secretion in the nasal mucosa does not prove increased expression in the epithelium in the lower airways, taken together the above data strongly suggest that cytokine elaboration is increased in the RV-infected lower airway epithelium.RV infection also induces cytokine production and adhesion molecule expression in cell culture. Divergent findings regard-

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A Stable HeLa Cell Line That Inducibly Expresses Poliovirus 2A ^pro : Effects on Cellular and Viral Gene Expression

Cited by 34 publications

References 71 publications

Differential Cleavage of eIF4GI and eIF4GII in Mammalian Cells

Differential Cleavage of eIF4GI and eIF4GII in Mammalian Cells

Proteinase 2A^proIs Essential for Enterovirus Replication in Type I Interferon-Treated Cells

Rhinovirus 16 3C Protease Induces Interleukin-8 and Granulocyte-Macrophage Colony-Stimulating Factor Expression in Human Bronchial Epithelial Cells

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A Stable HeLa Cell Line That Inducibly Expresses Poliovirus 2A pro : Effects on Cellular and Viral Gene Expression

Cited by 34 publications

References 71 publications

Differential Cleavage of eIF4GI and eIF4GII in Mammalian Cells

Differential Cleavage of eIF4GI and eIF4GII in Mammalian Cells

Proteinase 2AproIs Essential for Enterovirus Replication in Type I Interferon-Treated Cells

Rhinovirus 16 3C Protease Induces Interleukin-8 and Granulocyte-Macrophage Colony-Stimulating Factor Expression in Human Bronchial Epithelial Cells

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Product

Resources

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A Stable HeLa Cell Line That Inducibly Expresses Poliovirus 2A ^pro : Effects on Cellular and Viral Gene Expression

Proteinase 2A^proIs Essential for Enterovirus Replication in Type I Interferon-Treated Cells