Mapping of the Feline Calicivirus Proteinase Responsible for Autocatalytic Processing of the Nonstructural Polyprotein and Identification of a Stable Proteinase-Polymerase Precursor Protein

Sosnovtseva, Svetlana A.; Sosnovtsev, Stanislav V.; Green, Kim Y.

doi:10.1128/jvi.73.8.6626-6633.1999

Cited by 71 publications

(32 citation statements)

References 41 publications

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“…In some cases, the N-and C-terminal sites are cleaved with different kinetics. Thus, for example, C-terminal 3C/3CL pro cleavage occurs more slowly (picornaviruses) (36), is tightly regulated (arteriviruses) (45), or is totally lacking (some caliciviruses) (39,46). In our experiments, no evidence was obtained for cleavage in the region immediately downstream of the GAV 3CL pro which, according to comparative sequence analysis ( Fig.…”

Section: Discussioncontrasting

confidence: 53%

The 3C-Like Proteinase of an Invertebrate Nidovirus Links Coronavirus and Potyvirus Homologs

Ziebuhr

Bayer

Cowley

et al. 2003

J Virol

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Gill-associated virus (GAV), a positive-stranded RNA virus of prawns, is the prototype of newly recognized taxa (genus Okavirus, family Roniviridae) within the order Nidovirales. In this study, a putative GAV cysteine proteinase (3C-like proteinase [3CL pro ]), which is predicted to be the key enzyme involved in processing of the GAV replicase polyprotein precursors, pp1a and pp1ab, was characterized. Comparative sequence analysis indicated that, like its coronavirus homologs, 3CLpro has a three-domain organization and is flanked by hydrophobic domains. The putative 3CL pro domain including flanking regions (pp1a residues 2793 to 3143) was fused to the Escherichia coli maltose-binding protein (MBP) and, when expressed in E. coli, was found to possess N-terminal autoprocessing activity that was not dependent on the presence of the 3CL pro C-terminal domain. N-terminal sequence analysis of the processed protein revealed that cleavage occurred at the location 2827 LVTHE2VRTGN2836 . The trans-processing activity of the purified recombinant 3CL pro (pp1a residues 2832 to 3126) was used to identify another cleavage site, 6441 KVNHE2LYHVA 6450 , in the C-terminal pp1ab region. Taken together, the data tentatively identify VxHE2(L,V) as the substrate consensus sequence for the GAV 3CLpro . The study revealed that the GAV and potyvirus 3CL pro s possess similar substrate specificities which correlate with structural similarities in their respective substrate-binding sites, identified in sequence comparisons. Analysis of the proteolytic activities of MBP-3CL pro fusion proteins carrying replacements of putative active-site residues provided evidence that, in contrast to most other 3C/3CL pro s but in common with coronavirus 3CL pro s, the GAV 3CL pro employs a Cys 2968 -His 2879 catalytic dyad. The properties of the GAV 3CL pro define a novel RNA virus proteinase variant that bridges the gap between the distantly related chymotrypsin-like cysteine proteinases of coronaviruses and potyviruses.

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Section: Discussioncontrasting

confidence: 53%

The 3C-Like Proteinase of an Invertebrate Nidovirus Links Coronavirus and Potyvirus Homologs

Ziebuhr

Bayer

Cowley

et al. 2003

J Virol

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“…However, in another human pathogenic calicivirus, the sapovirus (SV), NS6 pro NS7 pol is the only protein detected in a cell-free system (Oka et al, 2005a,b). In the case of the pathogenic feline calicivirus (FCV), the NS6 pro NS7 pol precursor is detected in mammalian cells (Sosnovtsev et al, 2002(Sosnovtsev et al, , 1998Sosnovtseva et al, 1999). In accordance with our observations (Scheffler et al, 2007), evidence for autocatalytic cleavage of norovirus NS6 pro from the polyprotein precursor has been presented Liu et al, 1999;Seah et al, 1999;Sosnovtsev et al, 2006).…”

Section: The Viral Chymotrypsin-like Proteasesupporting

confidence: 87%

“…FCV has been used during the past two decades as a surrogate to investigate the replication strategy of the Caliciviridae. Many studies have brought insights into the replication of the vesivirus genome Green, 1995, 2000;Sosnovtsev et al, 2005Sosnovtsev et al, , 2003Sosnovtsev et al, , 1998Sosnovtseva et al, 1999). Lately, additional knowledge has been obtained from studies on the murine norovirus in RAW264.7 murine macrophages in vitro Wobus et al, 2004Wobus et al, , 2006.…”

Section: Multiplication Cycle Of the Caliciviridaementioning

confidence: 99%

Antiviral strategies to control calicivirus infections

et al. 2010

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Caliciviridae are human or non-human pathogenic viruses with a high diversity. Some members of the Caliciviridae, i.e. human pathogenic norovirus or rabbit hemorrhagic disease virus (RHDV), are worldwide emerging pathogens. The norovirus is the major cause of viral gastroenteritis worldwide, accounting for about 85% of the outbreaks in Europe between 1995 and 2000. In the United States, 25 million cases of infection are reported each year. Since its emergence in 1984 as an agent of fatal hemorrhagic diseases in rabbits, RHDV has killed millions of rabbits and has been dispersed to all of the inhabitable continents. In view of their successful and apparently increasing emergence, the development of antiviral strategies to control infections due to these viral pathogens has now become an important issue in medicine and veterinary medicine. Antiviral strategies have to be based on an understanding of the epidemiology, transmission, clinical symptoms, viral replication and immunity to infection resulting from infection by these viruses. Here, we provide an overview of the mechanisms underlying calicivirus infection, focusing on the molecular aspects of replication in the host cell. Recent experimental data generated through an international collaboration on structural biology, virology and drug design within the European consortium VIZIER is also presented. Based on this analysis, we propose antiviral strategies that may significantly impact on the epidemiological characteristics of these highly successful viral pathogens.

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“…ProPol still exhibited full protease activity and had consistently higher polymerase activity and for longer periods of time than Pol (46,47), suggesting that the ProPol form of the polymerase would be preferred by the virus. The same observation was made for the ProPol protein of feline calicivirus (FCV), another member of the Caliciviridae, in which ProPol was a predominant form of the RdRp observed in FCV-infected cells (48,49). Assuming that, like in the GI, both Pol and ProPol polymerase forms also exist in the GII.4 viruses, it is possible that the lower rate of polyprotein cleavage by the HOV Pro will result in the uncleaved or partially cleaved polyprotein components, among them ProPol, being present for longer periods of time, allowing the ProPol to more efficiently replicate viral RNA.…”

Section: Discussionmentioning

confidence: 53%

GII.4 Norovirus Protease Shows pH-Sensitive Proteolysis with a Unique Arg-His Pairing in the Catalytic Site

Viskovska

Zhao

Shanker

et al. 2019

J Virol

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Human noroviruses (NoVs) are the main cause of epidemic and sporadic gastroenteritis. Phylogenetically, noroviruses are divided into seven genogroups, with each divided into multiple genotypes. NoVs belonging to genogroup II and genotype 4 (GII.4) are globally most prevalent. Genetic diversity among the NoVs and the periodic emergence of novel strains present a challenge for the development of vaccines and antivirals to treat NoV infection. NoV protease is essential for viral replication and is an attractive target for the development of antivirals. The available structure of GI.1 protease provided a basis for the design of inhibitors targeting the active site of the protease. These inhibitors, although potent against the GI proteases, poorly inhibit the GII proteases, for which structural information is lacking. To elucidate the structural basis for this difference in the inhibitor efficiency, we determined the crystal structure of a GII.4 protease. The structure revealed significant changes in the S2 substratebinding pocket, making it noticeably smaller, and in the active site, with the catalytic triad residues showing conformational changes. Furthermore, a conserved arginine is found inserted into the active site, interacting with the catalytic histidine and restricting substrate/inhibitor access to the S2 pocket. This interaction alters the relationships between the catalytic residues and may allow for a pHdependent regulation of protease activity. The changes we observed in the GII.4 protease structure may explain the reduced potency of the GI-specific inhibitors against the GII protease and therefore must be taken into account when designing broadly cross-reactive antivirals against NoVs. IMPORTANCE Human noroviruses (NoVs) cause sporadic and epidemic gastroenteritis worldwide. They are divided into seven genogroups (GI to GVII), with each genogroup further divided into several genotypes. Human NoVs belonging to genogroup II and genotype 4 (GII.4) are the most prevalent. Currently, there are no vaccines or antiviral drugs available for NoV infection. The protease encoded by NoV is considered a valuable target because of its essential role in replication. NoV protease structures have only been determined for the GI genogroup. We show here that the structure of the GII.4 protease exhibits several significant changes from GI proteases, including a unique pairing of an arginine with the catalytic histidine that makes the proteolytic activity of GII.4 protease pH sensitive. A comparative analysis of NoV protease structures may provide a rational framework for structure-based drug design of broadly cross-reactive inhibitors targeting NoVs.

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Mapping of the Feline Calicivirus Proteinase Responsible for Autocatalytic Processing of the Nonstructural Polyprotein and Identification of a Stable Proteinase-Polymerase Precursor Protein

Cited by 71 publications

References 41 publications

The 3C-Like Proteinase of an Invertebrate Nidovirus Links Coronavirus and Potyvirus Homologs

The 3C-Like Proteinase of an Invertebrate Nidovirus Links Coronavirus and Potyvirus Homologs

Antiviral strategies to control calicivirus infections

GII.4 Norovirus Protease Shows pH-Sensitive Proteolysis with a Unique Arg-His Pairing in the Catalytic Site

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