Cytoplasmic redistribution and cleavage of AUF1 during coxsackievirus infection enhance the stability of its viral genome

Wong, Jerry; Si, Xiaoning; Angeles, Arkhjamil; Zhang, Jingchun; Shi, Junyan; Gabriel, Franck; Jagdeo, Julienne; Wang, Tianying; Zhong, Zhaohua; Jan, Eric; Luo, Honglin

doi:10.1096/fj.12-226498

Cited by 41 publications

(57 citation statements)

References 51 publications

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“…Among them, several members of the hnRNPs family have been previously shown to play a role in regulating viral replication. 7,8,39,[44][45][46][47] For examples, hnRNP C and K were documented to interact with the non-coding regions of enteroviral RNA to facilitate translation initiation and RNA synthesis. 44,45,47 Similar to our findings in the current study, enteroviral infection causes cytoplasmic redistribution and cleavage of hnRNP D and deletion of this protein results in enhanced viral replication, suggesting that hnRNA D acts as a host restriction factor against enterovirus infection.…”

Section: Discussionmentioning

confidence: 99%

“…44,45,47 Similar to our findings in the current study, enteroviral infection causes cytoplasmic redistribution and cleavage of hnRNP D and deletion of this protein results in enhanced viral replication, suggesting that hnRNA D acts as a host restriction factor against enterovirus infection. 7,8,39 The antiviral mechanisms of hnRNP D appear to involve negative regulation of internal ribosome entry site (IRES)-mediated translation of viral RNA via direct interaction with the IRES of enteroviruses, 7,46 and viral RNA degradation by targeting the 3′-untranslated region (UTR) of enteroviral genome. 39 The exact mechanism of TDP-43 suppressing CVB3 replication remains to be clarified.…”

Section: Discussionmentioning

confidence: 99%

“…7,8,39 The antiviral mechanisms of hnRNP D appear to involve negative regulation of internal ribosome entry site (IRES)-mediated translation of viral RNA via direct interaction with the IRES of enteroviruses, 7,46 and viral RNA degradation by targeting the 3′-untranslated region (UTR) of enteroviral genome. 39 The exact mechanism of TDP-43 suppressing CVB3 replication remains to be clarified. In this study, we tested our hypothesis that TDP-43 interacts directly with the viral RNA through its RRM domains, leading to the inhibition of viral translation and RNA replication.…”

Section: Discussionmentioning

confidence: 99%

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Cytoplasmic translocation, aggregation, and cleavage of TDP-43 by enteroviral proteases modulate viral pathogenesis

et al. 2015

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We have previously demonstrated that infection by coxsackievirus B3 (CVB3), a positive-stranded RNA enterovirus, results in the accumulation of insoluble ubiquitin-protein aggregates, which resembles the common feature of neurodegenerative diseases. The importance of protein aggregation in viral pathogenesis has been recognized; however, the underlying regulatory mechanisms remain ill-defined. Transactive response DNA-binding protein-43 (TDP-43) is an RNA-binding protein that has an essential role in regulating RNA metabolism at multiple levels. Cleavage and cytoplasmic aggregation of TDP-43 serves as a major molecular marker for amyotrophic lateral sclerosis and frontotemporal lobar degeneration and contributes significantly to disease progression. In this study, we reported that TDP-43 is translocated from the nucleus to the cytoplasm during CVB3 infection through the activity of viral protease 2A, followed by the cleavage mediated by viral protease 3C. Cytoplasmic translocation of TDP-43 is accompanied by reduced solubility and increased formation of protein aggregates. The cleavage takes place at aminoacid 327 between glutamine and alanine, resulting in the generation of an N-and C-terminal cleavage fragment of~35 and~8 kDa, respectively. The C-terminal product of TDP-43 is unstable and quickly degraded through the proteasome degradation pathway, whereas the N-terminal truncation of TDP-43 acts as a dominant-negative mutant that inhibits the function of native TDP-43 in alternative RNA splicing. Lastly, we demonstrated that knockdown of TDP-43 results in an increase in viral titers, suggesting a protective role for TDP-43 in CVB3 infection. Taken together, our findings suggest a novel model by which cytoplasmic redistribution and cleavage of TDP-43 as a consequence of CVB3 infection disrupts the solubility and transcriptional activity of TDP-43. Our results also reveal a mechanism evolved by enteroviruses to support efficient viral infection. Coxsackievirus B3 (CVB3) is a small, positive-stranded RNA enterovirus. 1 The single open reading frame of CVB3 is translated into a viral polypeptide that is subsequently cleaved by two virus-encoded proteases 2A and 3C to generate structural and non-structural proteins. 2 In addition to processing viral polyprotein, 2A and 3C target host proteins important for maintenance of protein translation and transcription, antiviral activity, and cellular architecture and signaling, contributing to virus-induced pathogenesis. [3][4][5] Although enteroviral replication takes place exclusively in the cytoplasm, viral infection has been demonstrated to lead to cytoplasmic translocation of nuclear proteins. 6 For example, heterogeneous ribonucleoprotein D (hnRNP D) has been shown to translocate from the nucleus to the cytoplasm during enteroviral infection. 5,7,8 Moreover, hnRNP D is cleaved by 3C and has an antiviral function against enteroviral infection. 5,7,8 Cytoplasmic translocation after enteroviral infection has also been demonstrated for several other hnRNPs (A1, C, and K)...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Cytoplasmic translocation, aggregation, and cleavage of TDP-43 by enteroviral proteases modulate viral pathogenesis

et al. 2015

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“…In addition to NBR1 and SQSTM1, CVB3 proteases were reported to target a number of other host proteins, such as eukaryotic translation initiation factor 4g, 32 poly(A)-binding protein, 33 serum response factor, 34 dystrophin, 35 adenosineuridine-rich element RNA binding factor 1, 36 GTPase activating protein (SH3 domain) binding protein 1, 37 melanoma differentiation-associated protein 5, 38 mitochondrial antiviralsignaling protein 38 and retinoic acid-inducible gene 1.…”

Section: Discussionmentioning

confidence: 99%

Dominant-negative function of the C-terminal fragments of NBR1 and SQSTM1 generated during enteroviral infection

et al. 2014

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Coxsackievirus infection induces an abnormal accumulation of ubiquitin aggregates that are generally believed to be noxious to the cells and have a key role in viral pathogenesis. Selective autophagy mediated by autophagy adaptor proteins, including sequestosome 1 (SQSTM1/p62) and neighbor of BRCA1 gene 1 protein (NBR1), are an important pathway for disposing of misfolded/ubiquitin conjugates. We have recently demonstrated that SQSTM1 is cleaved after coxsackievirus infection, resulting in the disruption of SQSTM1 function in selective autophagy. NBR1 is a functional homolog of SQSTM1. In this study, we propose to test whether NBR1 can compensate for the compromise of SQSTM1 after viral infection. Of interest, we found that NBR1 was also cleaved after coxsackievirus infection. This cleavage took place at two sites mediated by virus-encoded protease 2A pro and 3C pro , respectively. In addition to the loss-of-function, we further investigated whether cleavage of SQSTM1/NBR1 leads to the generation of toxic gain-of-function mutants. We showed that the C-terminal fragments of SQSTM1 and NBR1 exhibited a dominant-negative effect against native SQSTM1/NBR1, probably by competing for LC3 and ubiquitin chain binding. Finally, we demonstrated a positive, mutual regulatory relationship between SQSTM1 and NBR1 during viral infection. We showed that knockdown of SQSTM1 resulted in reduced expression of NBR1, whereas overexpression of SQSTM1 led to increased level of NBR1, and vice versa, further excluding the possible compensation of NBR1 for the loss of SQSTM1. Taken together, the findings in this study suggest a novel mechanism through which coxsackievirus infection induces increased accumulation of ubiquitin conjugates and subsequent viral damage.

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“…roteases play fundamental roles in cells by ultimately changing the fate and function of its substrates through proteolytic cleavage or degradation (1)(2)(3)(4)(5)(6)(7)(8)(9)72). Many viruses have exploited these strategies to promote infection.…”

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confidence: 99%

Heterogeneous Nuclear Ribonucleoprotein M Facilitates Enterovirus Infection

Jagdeo

Dufour

Gabriel

et al. 2015

J Virol

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Picornavirus infection involves a dynamic interplay of host and viral protein interactions that modulates cellular processes to facilitate virus infection and evade host antiviral defenses. Here, using a proteomics-based approach known as TAILS to identify protease-generated neo-N-terminal peptides, we identify a novel target of the poliovirus 3C proteinase, the heterogeneous nuclear ribonucleoprotein M (hnRNP M), a nucleocytoplasmic shuttling RNA-binding protein that is primarily known for its role in pre-mRNA splicing. hnRNP M is cleaved in vitro by poliovirus and coxsackievirus B3 (CVB3) 3C proteinases and is targeted in poliovirus-and CVB3-infected HeLa cells and in the hearts of CVB3-infected mice. hnRNP M relocalizes from the nucleus to the cytoplasm during poliovirus infection. Finally, depletion of hnRNP M using small interfering RNA knockdown approaches decreases poliovirus and CVB3 infections in HeLa cells and does not affect poliovirus internal ribosome entry site translation and viral RNA stability. We propose that cleavage of and subverting the function of hnRNP M is a general strategy utilized by picornaviruses to facilitate viral infection. IMPORTANCEEnteroviruses, a member of the picornavirus family, are RNA viruses that cause a range of diseases, including respiratory ailments, dilated cardiomyopathy, and paralysis. Although enteroviruses have been studied for several decades, the molecular basis of infection and the pathogenic mechanisms leading to disease are still poorly understood. Here, we identify hnRNP M as a novel target of a viral proteinase. We demonstrate that the virus subverts the function of hnRNP M and redirects it to a step in the viral life cycle. We propose that cleavage of hnRNP M is a general strategy that picornaviruses use to facilitate infection. P roteases play fundamental roles in cells by ultimately changing the fate and function of its substrates through proteolytic cleavage or degradation (1-9, 72). Many viruses have exploited these strategies to promote infection. For instance, picornaviruses translate their RNA genome as a single polyprotein, which is then processed by virus-encoded proteinases to produce the mature viral proteins. In addition, viral proteinases target a subset of host proteins to modulate cellular processes, inhibit antiviral responses, and subvert host proteins to aid in specific steps of the viral life cycle, such as viral translation and replication. One of the best characterized is the rapid host translational shutoff that occurs in poliovirus-infected cells, which is facilitated through cleavage of eukaryotic translation initiation factors eIF4G and the poly(A) binding protein (PABP) by the viral proteinases 2A and 3C (10-14). Overall inhibition of protein synthesis leads to release of translation factors and ribosomes that are then diverted to poliovirus protein synthesis. The identification of host protein substrates of viral proteinases has provided important insights into the virus-host interaction strategies that contribute to the...

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Cytoplasmic redistribution and cleavage of AUF1 during coxsackievirus infection enhance the stability of its viral genome

Cited by 41 publications

References 51 publications

Cytoplasmic translocation, aggregation, and cleavage of TDP-43 by enteroviral proteases modulate viral pathogenesis

Cytoplasmic translocation, aggregation, and cleavage of TDP-43 by enteroviral proteases modulate viral pathogenesis

Dominant-negative function of the C-terminal fragments of NBR1 and SQSTM1 generated during enteroviral infection

Heterogeneous Nuclear Ribonucleoprotein M Facilitates Enterovirus Infection

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