The Viral Nucleocapsid Protein of Transmissible Gastroenteritis Coronavirus (TGEV) Is Cleaved by Caspase-6 and -7 during TGEV-Induced Apoptosis

Éléouët, Jean-François; Slee, Elizabeth A.; Saurini, Françoise; Castagné, Nathalie; Poncet, Didier; Garrido, Carmen; Solary, Éric; Martin, Séamus J.

doi:10.1128/jvi.74.9.3975-3983.2000

Cited by 88 publications

(81 citation statements)

References 46 publications

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“…It has been demonstrated that host cell caspases, which are activated during coronavirus infection, are responsible for this cleavage (31). A common caspase cleavage motif is present in all of the mentioned coronavirus N proteins.…”

Section: Discussionmentioning

confidence: 99%

Mass Spectrometric Characterization of Proteins from the SARS Virus

Krokhin

Li²,

Andonov³

et al. 2003

Molecular & Cellular Proteomics

162

168

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A new coronavirus has been implicated as the causative agent of severe acute respiratory syndrome (SARS). We have used convalescent sera from several SARS patients to detect proteins in the culture supernatants from cells exposed to lavage another SARS patient. The most prominent protein in the supernatant was identified by matrixassisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as a ϳ46-kDa species. This was found to be a novel nucleocapsid protein that matched almost exactly one predicted by an open reading frame in the recently published nucleotide sequence of the same virus isolate (>96% coverage). A second viral protein corresponding to the predicted ϳ139-kDa spike glycoprotein has also been examined by MALDI-TOF MS (42% coverage). After peptide N-glycosidase F digestion, 12 glycosylation sites in this protein were confirmed. The sugars attached to four of the sites were also identified. These results suggest that the nucleocapsid protein is a major immunogen that may be useful for early diagnostics, and that the spike glycoprotein may present a particularly attractive target for prophylactic intervention in combating SARS. Molecular & Cellular Proteomics 2: 346 -356, 2003.

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Section: Discussionmentioning

confidence: 99%

Mass Spectrometric Characterization of Proteins from the SARS Virus

Krokhin

Li²,

Andonov³

et al. 2003

Molecular & Cellular Proteomics

162

168

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“…During transmissible gastroenteritis coronavirus infection, caspase activation causes cleavage of the viral nucleocapsid protein (26). During infection with influenza, virus cleavage of the nucleocapsid protein has been observed during the process of apoptosis in late stages of the infection, and the primary structure of NP of human influenza viruses (A and B) has been shown to have proteolytic sites, which engage involvement of caspases (27).…”

Section: Discussionmentioning

confidence: 99%

Induction of Caspase Activation and Cleavage of the Viral Nucleocapsid Protein in Different Cell Types during Crimean-Congo Hemorrhagic Fever Virus Infection

Karlberg

Tan

Mirazimi

2011

Journal of Biological Chemistry

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Regulation of apoptosis during infection has been observed for several viral pathogens. Programmed cell death and regulation of apoptosis in response to a viral infection are important factors for host or virus survival. It is not known whether Crimean-Congo hemorrhagic fever virus (CCHFV) infection regulates the apoptosis process in vitro. This study for the first time suggests that CCHFV induces apoptosis, which may be dependent on caspase-3 activation. This study also shows that the coding sequence of the S segment of CCHFV contains a proteolytic cleavage site, DEVD, which is conserved in all CCHFV strains. By using different recombinant expression systems and site-directed mutagenesis, we demonstrated that this motif is subject to caspase cleavage. We also demonstrate that CCHFV nucleocapsid protein (NP) is cleaved into a 30-kDa fragment at the same time as caspase activity is induced during infection. Using caspase inhibitors and cells lacking caspase-3, we clearly demonstrate that the cleavage of NP is caspase-3-dependent. We also show that the inhibition of apoptosis induced progeny viral titers of ϳ80 -90%. Thus, caspase-3-dependent cleavage of NP may represent a host defense mechanism against lytic CCHFV infection. Taken together, these data suggest that the most abundant protein of CCHFV, which has several essential functions such as protection of viral RNA and participation in various processes in the replication cycle, can be subjected to cleavage by host cell caspases.Programmed cell death and regulation of apoptosis in response to a viral infection are important factors for host or virus survival. Protease caspases (cysteine aspartate-specific proteases) play an important role in apoptosis. Caspase activation leads to a proteolytic cascade, where procaspase-8, -9, -10 is activated. These initiator caspases activate caspase-3, -6, and -7, which organize the death of the cell. Caspase-3 is activated as a mediator in the effector phase of programmed cell death. The activated caspases cleave the substrates at specific sites, making them very specific members of the proteases (1). Poly(ADP-ribose) polymerase (PARP) 2 is one substrate responsible for the destruction of cellular structures (2, 3). It has been demonstrated that PARP can be cleaved by activated caspase-3 and -7.To date, several studies have demonstrated the regulation of apoptosis in the replication cycle and spread of viruses. Epstein-Barr virus encodes proteins for inhibition of different steps of the apoptotic pathway in order to maintain virus persistence (4). In contrast to Epstein-Barr virus, vesicular stomatitis virus (VSV) induces cell death. Induced apoptosis during VSV infection is triggered at an early stage of infection and does not depend on viral protein synthesis (5). Intravascular apoptosis and destruction of immune cells have also been observed during infection in fatal cases of Ebola virus. However, the mechanism behind this event is not fully known.The family Bunyaviridae is one of the largest virus groups, comprising o...

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“…On the other hand, however, it is now well recognized that caspase-mediated cleavage of many viral proteins is a direct requirement for propagation of those viruses (10). Specifically, caspase 2 has been implicated in the cleavage of FCV capsid (14), whereas caspase 6 was found to be involved in the processing of human astrovirus HAstV capsid precursor (12), in TGEV nucleocapsid cleavage (11), and in HIV-induced apoptosis of T lymphocytes (103). Inasmuch as limited cleavage of proteins can either lead to the functional inhibition or activation of these proteins (104 -105) we could hypothesize that caspase-mediated cleavage of the NS5A protein may modulate the multiple functions of the protein.…”

Section: Discussionmentioning

confidence: 99%

“…Several caspases have been shown to target the structural proteins of different viruses, including transmissible gastroenteritis coronavirus (TGEV) (11), human astrovirus (HAstv) (12), influenza A virus (13), and feline calicivirus (FCV) (14), as well as nonstructural viral proteins, such as the NS1 from the Aleutian mink disease parvovirus (ADV) (15), the adenovirus (Ad) E1A (16), or the immediately early protein 22 from herpes simplex virus (HSV-1) (17), with an impact on viral pathogenesis. Interestingly, caspase 3, the executioner caspase that targets the majority of cellular substrates during apoptosis, is also responsible for cleaving many viral proteins, and numerous studies have implicated caspase 6, 7, and 2 in the cleavage of different viral protein substrates (11,14,16). Because most viruses encode inhibitors of caspases to evade cellular antiviral mechanisms that lead cells to apoptosis, (18 -21) it is not immediately apparent why viruses rely on caspases for cleavage of their own proteins.…”

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confidence: 99%

The NS5A Protein of the Hepatitis C Virus Genotype 1a Is Cleaved by Caspases to Produce C-terminal-truncated Forms of the Protein That Reside Mainly in the Cytosol

Kalamvoki

Georgopoulou

Mavromara

2006

Journal of Biological Chemistry

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The nonstructural 5A (NS5A) protein of the hepatitis C virus (HCV) is a multifunctional protein that is implicated in viral replication and pathogenesis. We report here that NS5A of HCV-1a is cleaved at multiple sites by caspase proteases in transfected cells. Two cleavage sites at positions Asp 154 and 248 DXXD 251 were mapped. Cleavage at Asp 154 has been previously recognized as one of the caspase cleavage sites for the NS5A protein of HCV genotype 1b (1, 2) and results in the production of a 17-kDa fragment. The sequence 248 DXXD 251 is a novel caspase recognition motif for NS5A and is responsible for the production of a 31-kDa fragment. Furthermore, we show that Arg 217 is implicated in the production of the previously described 24-kDa product, whose accumulation is affected by both calpain and caspase inhibitors. We also showed that caspase-mediated cleavage occurs in the absence of exogenous proapoptotic stimuli and is not related to the accumulation of the protein in the endoplasmic reticulum. Interestingly, our data indicate that NS5A is targeted by at least two different caspases and suggest that caspase 6 is implicated in the production of the 17-kDa fragment. Most importantly, we report that, all the detectable NS5A fragments following caspase-mediated cleavage are C-terminaltruncated forms of NS5A and are mainly localized in the cytosol. Thus, in sharp contrast to the current view we found no evidence supporting a role for caspase-mediated cleavage in the transport of the NS5A protein to the nucleus, which could lead to transcriptional activation.

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The Viral Nucleocapsid Protein of Transmissible Gastroenteritis Coronavirus (TGEV) Is Cleaved by Caspase-6 and -7 during TGEV-Induced Apoptosis

Cited by 88 publications

References 46 publications

Mass Spectrometric Characterization of Proteins from the SARS Virus

Mass Spectrometric Characterization of Proteins from the SARS Virus

Induction of Caspase Activation and Cleavage of the Viral Nucleocapsid Protein in Different Cell Types during Crimean-Congo Hemorrhagic Fever Virus Infection

The NS5A Protein of the Hepatitis C Virus Genotype 1a Is Cleaved by Caspases to Produce C-terminal-truncated Forms of the Protein That Reside Mainly in the Cytosol

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