The coding region for the 54-kDa protein of several pestiviruses lacks host insertions but reveals a “Zinc finger-like” domain

Moerlooze, Laurence De; Desport, Moira; Renard, A.; Lecomte, C.; Brownlie, Joe; Martial, Joseph

doi:10.1016/0042-6822(90)90555-6

Cited by 35 publications

(22 citation statements)

References 22 publications

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“…The upstream insertion of 30 aa includes five Cys and one His that are conserved among all known pestivirus isolates; these residues are organized in a linear fashion, suggesting the presence of a mononuclear ZnB site (12). Accordingly, two amino acid pairs may coordinate a Zn 2ϩ ion: candidate members for the N-terminal pair are H1483 or C1484, and C1487, and C1490, while C1500 and C1503 may form the C-terminal pair (Fig.…”

Section: Resultsmentioning

confidence: 99%

Temporal Modulation of an Autoprotease Is Crucial for Replication and Pathogenicity of an RNA Virus

Lackner

Müller

Pankraz

et al. 2004

J Virol

126

196

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Pestiviruses belong to the family Flaviviridae, and their genome is a single-stranded RNA of positive polarity encoding one large polyprotein which is further processed into mature proteins. Noncytopathogenic (noncp) strains of the pestivirus bovine viral diarrhea virus (BVDV) can establish persistent infection. In persistently infected animals, noncp BVDVs occasionally acquire mutations in viral nonstructural protein 2 (NS2) that give rise to cytopathogenic (cp) BVDV variants, and, eventually, lead to the onset of lethal disease. A molecular marker of cp BVDV infection is a high-level expression of the replicative NS3 protease/helicase that together with NS2 is derived from NS2-3. Here, we present evidence for NS2-3 autoprocessing by a newly identified cysteine protease in NS2 that is distantly related to the NS2-3 autoprotease of hepatitis C and GB viruses. The vital role of this autoprotease in BVDV infection was established, implying an essential function for NS3 in pestiviral RNA replication which cannot be supplied by its NS2-3 precursor. Accordingly, and contrary to a current paradigm, we detected almost complete cleavage of NS2-3 in noncp BVDV at early hours of infection. At 6 to 9 h postinfection, NS2-3 autoprocessing diminished to barely detectable levels for noncp BVDV but decreased only moderately for cp BVDV. Viral RNA synthesis rates strictly correlated with different NS3 levels in noncp and cp BVDV-infected cells, implicating the NS2 autoprotease in RNA replication control. The biotype-specific modulation of NS2-3 autoprocessing indicates a crucial role of the NS2 autoprotease in the pathogenicity of BVDV.Pestiviruses are animal pathogens that are recognized as a separate genus of the family Flaviviridae, which also includes the genera Flavivirus and Hepacivirus (hepatitis C viruses [HCV]), as well as the unassigned GB viruses (32). Pestiviruses are widely used as a surrogate model for studying HCV, which grows poorly in available cell culture systems. Persistent HCV infections are a major cause of liver cirrhosis and hepatocellular carcinoma in humans worldwide.The pestiviral genome is a positive-stranded RNA of 12.3 kb. It is translated into a large polyprotein, which is cotranslationally and posttranslationally processed by viral and cellular proteases. The order of proteins in the polyprotein is NH 2 -N pro -C-E rns -E1-E2-p7-NS2-3-NS4A-NS4B-NS5A-NS5B-COOH. The autoprotease N pro generates its C terminus and the N terminus of the downstream core protein C. The proteolytic releases of the structural glycoproteins E rns (RNase secreted), E1, E2, and p7 are mediated by cellular signal peptidases. The nonstructural protein 4A (NS4A)-dependent chymotrypsin-like serine protease in NS3 mediates processing in the NS region downstream of NS3 (32). The mechanism of NS2-3 cleavage was hitherto unknown and is the subject of this study (see below).The pestivirus bovine viral diarrhea virus (BVDV) can establish lifelong persistent infections in animals, which become the primary sources for the horizontal sp...

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Section: Resultsmentioning

confidence: 99%

Temporal Modulation of an Autoprotease Is Crucial for Replication and Pathogenicity of an RNA Virus

Lackner

Müller

Pankraz

et al. 2004

J Virol

126

196

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“…gp48 gp25 gp53 p54 p80 p58 p75 detailed in a previous report (Lecomte et al, 1990). All the RNAs led to specific PCR products of the correct size.…”

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confidence: 99%

Nucleotide sequence of the bovine viral diarrhoea virus Osloss strain: comparison with related viruses and identification of specific DNA probes in the 5' untranslated region

Moerlooze

Lecomte

Brown-Shimmer

et al. 1993

Journal of General Virology

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“…This hypothesis is supported paradoxically by our result that CP BVD viruses change to NCP biotype by deletion of the cellular mRNA integrated in the region encoding p125 nonstructural protein. The existence of foreign RNA insertion into the portion of the genome encoding for the p125 polypeptide have been reported for several, but not all, CP BVD viruses [7,9,12,[20][21][22][23]25]. Therefore, the occurrence of insertion may not be the only mechanism involved in biotypic determination [27].…”

Section: Discussionmentioning

confidence: 99%

“…This hypothesis, however, was challenged by observations that some CP BVD viruses lacked the insertion in their genomes [7,9,12]. Therefore, the understanding of the cytopathogenicity of the BVD virus seems far from complete.…”

mentioning

confidence: 93%

“…proposed that a NCP BVD virus mutated to a CP BVD virus by taking up cellular sequences during a recombination event in persistently infected animals [21,22]. This hypothesis, however, was challenged by observations that some CP BVD viruses lacked the insertion in their genomes [7,9,12]. Therefore, the understanding of the cytopathogenicity of the BVD virus seems far from complete.…”

mentioning

confidence: 99%

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Variation from Cytopathogenic Biotype to Non-Cytopathogenic Biotype is Correlated with the Deletion of Cellular Sequence from Bovine Viral Diarrhea Viruses.

Nakamura¹,

Sakamoto

Sakoda

et al. 1997

The Journal of Veterinary Medical Science

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ABSTRACT. Non-cytopathogenic (NCP) viruses of bovine viral diarrhea (BVD) virus were detected at a low ratio by the reverse plaque formation method from virus samples after several plaque clonings of cytopathogenic (CP) BVD viruses; NADL and Osloss strains. This phenomenon suggests that the NCP BVD viruses are produced at a low ratio during the propagation of CP BVD viruses in vitro. To investigate the differences between the parent CP BVD virus and the NCP BVD virus as a real progeny, the regions flanking the insertion of cellular mRNA in the p125 domain of NADL and Osloss strains were amplified by RT-PCR and cloned into pGEM 3Z plasmid vector, and then sequenced. Consequently, it was confirmed that sequences of cellular mRNA insertion of CP BVD viruses, NADL and Osloss strains, were completely and exactly deleted from the NCP BVD viruses which were real progeny of CP BVD viruses, NADL and Osloss strains. These results suggest that NCP BVD viruses may revert from CP biotype to NCP biotype by the deletion of cellular mRNA insertion in the viral genome of CP BVD viruses (NADL and Osloss strains). proposed that a NCP BVD virus mutated to a CP BVD virus by taking up cellular sequences during a recombination event in persistently infected animals [21,22]. This hypothesis, however, was challenged by observations that some CP BVD viruses lacked the insertion in their genomes [7,9,12]. Therefore, the understanding of the cytopathogenicity of the BVD virus seems far from complete. Recently, NCP BVD viruses were isolated from CP BVD virus stocks and each virus stock of CP BVD virus contained simultaneously two kinds of NCP BVD viruses showing a different manner of interference [24]. It was speculated that the mutation from the CP biotype to the NCP biotype occurred during the propagation of CP BVD viruses in vitro. This paper deals with the possibility that the reversion of BVD virus from CP biotype to NCP biotype is caused by the deletion of cellular mRNA insertion in viral genome of CP BVD viruses; NADL and Osloss strains. MATERIALS AND METHODS Viruses and cells:The NADL [13] and Osloss strains [26] of CP BVD virus and derivative NCP BVD viruses (NADL-END + , NADL-END -, Osloss-END + and Osloss-END -strains) newly isolated from plaque cloned NADL and Osloss strains in our laboratory were used in this study. The NADL strain originated from the American Type Culture Collection (Rockville, MD, U.S.A.), and the Osloss strain was kindly provided by Prof. B. Liess, Institute for Virology, Hannover Veterinary School, Germany. Stock viruses were propagated in bovine testicle (BT) cell cultures and stored at 80°C. The BT cells were grown in Eagle's minimum essential medium (MEM) containing 10% fetal

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The coding region for the 54-kDa protein of several pestiviruses lacks host insertions but reveals a “Zinc finger-like” domain

Cited by 35 publications

References 22 publications

Temporal Modulation of an Autoprotease Is Crucial for Replication and Pathogenicity of an RNA Virus

Temporal Modulation of an Autoprotease Is Crucial for Replication and Pathogenicity of an RNA Virus

Nucleotide sequence of the bovine viral diarrhoea virus Osloss strain: comparison with related viruses and identification of specific DNA probes in the 5' untranslated region

Variation from Cytopathogenic Biotype to Non-Cytopathogenic Biotype is Correlated with the Deletion of Cellular Sequence from Bovine Viral Diarrhea Viruses.

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