The genomic RNA of pestiviruses is translated into a large polyprotein that cleaved number proteins. structural proteins are N terminal in this and include three glycoproteins called E0, E1, E2 on the basis order which they appear polyprotein. Using pulse-chase experiments, we show pestiviral glycoprotein precursor, E012, formed processed an ordered fashion. Processing initiated by nascent cleavage between capsid translocated E012 followed at C terminus E2. then rapidly to form E01 After...

Virions from hog cholera virus (HCV), a member of the genus Pestivirus, were analyzed by using specific antibodies. The nucleocapsid protein was found to be 14-kDa molecule (HCV p14). An equivalent could also demonstrated for virions another pestivirus, bovine viral diarrhea virus. HCV envelope is composed three glycoproteins, gp44/48, gp33, and gp55. All exist in form disulfide-linked dimers virus-infected cells virions; gp44/48 gp55 each homodimers, whereas dimerized with gp33. Such...

Neutralizing monoclonal antibodies directed against hog cholera virus (HCV) precipitated two HCV-encoded glycoproteins, HCV gp55 and gp33. Immunoassay with bacterial fusion proteins Western immunoblotting extracts from infected cells revealed that the recognized only gp55. Coprecipitation of gp33 was shown to be due intermolecular disulfide bridges. One also reacted major glycoprotein another pestivirus, bovine viral diarrhea (BVDV). The analogous BVDV glycoproteins exhibited a distribution...

Upon infection of animal cells by Sindbis virus, four nonstructural (ns) proteins, termed nsP1-4 in order from 5' to 3' the genome, are produced posttranslational cleavage a polyprotein. nsP4 is believed function as viral RNA polymerase and short-lived infected cells. We show here that reticulocyte lysates degraded N-end rule pathway, one ubiquitin-dependent proteolytic pathway. When N-terminal residue changed mutagenesis, metabolic stabilities mutant nsP4s follow rule, half-life bearing...

A cDNA fragment covering the genomic region that encodes structural proteins of hog cholera virus (HCV) was inserted into tk gene vaccinia virus. Expression studies with virus/HCV recombinants led to identification HCV-specific proteins. The putative HCV core protein p23 demonstrated for first time by using an antiserum against a bacterial fusion protein. glycoproteins expressed recombinant migrated on sodium dodecyl sulfate-gels identically precipitated from HCV-infected cells....

In 2013, several Austrian piglet-producing farms recorded outbreaks of action-related repetitive myoclonia in newborn piglets ("shaking piglets"). Malnutrition was seen numerous as a complication this tremor syndrome. Overall piglet mortality increased and the number weaned per sow decreased by more than 10% due to outbreak. Histological examination CNS affected revealed moderate hypomyelination white substance cerebellum spinal cord. We detected recently discovered pestivirus, termed...

Western equine encephalomyelitis (WEE) virus (Togaviridae: Alphavirus) was shown previously to have arisen by recombination between eastern (EEE)- and Sindbis-like viruses (C. S. Hahn, Lustig, E. G. Strauss, J. H. Proc. Natl. Acad. Sci. USA 85:5997-6001, 1988). We now examined the recombinational history evolution of all belonging WEE antigenic complex, including Buggy Creek, Fort Morgan, Highlands J, Sindbis, Babanki, Ockelbo, Kyzylagach, Whataroa, Aura viruses, using nucleotide sequences...

The polyprotein of classical swine fever virus starts with the nonstructural protein p23, which is followed by nucleocapsid p14. Proteolytic cleavage between p23 and p14 was demonstrated in a cell-free transcription-translation system. Successive truncation cDNA used for transcription indicated that proteolytic activity responsible resides within p23. In order to determine site these two proteins, respective genomic regions were expressed different expression systems. N-terminal sequencing...

The 5'- and 3'-terminal sequences of the genomic RNA from classical swine fever virus (CSFV) were determined, resulting information was used for construction full-length CSFV cDNA clones. After transfection in vitro-transcribed derived a construct, infectious recovered porcine cells. To confirm de novo generation cloned DNA, genetically tagged constructed. In comparison with parental CSFV, recombinant viruses retarded growth by about 1 order magnitude. Introduction deletion exchange part...

The pestivirus genome encodes a single polyprotein which is subject to co- and posttranslational processing by cellular viral proteases. map positions of all virus-encoded proteins are known with the exception hypothetical peptide (p?) interlinks glycoprotein E2 nonstructural protein NS2-3 approximately between amino acid 1060 1130. Expression studies recombinant vaccinia viruses bearing set C-terminally truncated E2-p?-NS2-encoding sequences derived from bovine diarrhea virus (BVDV) strain...

ABSTRACT Various monoclonal antibodies (MAbs) that recognize cell surface proteins on bovine cells were previously shown to efficiently block infection with viral diarrhea virus (BVDV) (C. Schelp, I. Greiser-Wilke, G. Wolf, M. Beer, V. Moennig, and B. Liess, Arch. Virol. 140: 1997-2009, 1995). With one of these MAbs, a 50- 58-kDa protein was purified from calf thymus by immunoaffinity chromatography. Microchemical analysis two internal peptides revealed significant sequence homology porcine...

Proteolytic processing of polyproteins is considered a crucial step in the life cycle most positive-strand RNA viruses. An enhancement NS2-3 has been described as major difference between noncytopathogenic (non-CP) and cytopathogenic (CP) biotypes pestiviruses. The effects accelerated versus delayed on maturation other nonstructural proteins (NSP) have never compared. In this study, we analyzed proteolytic NSP Classical swine fever virus (CSFV). Key to investigation was panel newly developed...

European honey bees are highly important in crop pollination, increasing the value of global agricultural production by billions dollars. Current knowledge about virulence and pathogenicity Deformed wing virus (DWV), a major factor bee colony mortality, is limited. With this study, we close gap between field research laboratory investigations establishing complete vitro model for DWV pathogenesis. Infectious was rescued from molecular clone DWV-A genome that induces symptoms such as crippled...

Abstract A novel pestivirus species was discovered in a piglet-producing farm Austria during virologic examinations of congenital tremor cases. The emergence this species, provisionally termed Linda virus, domestic pigs may have implications for classical swine fever virus surveillance and porcine health management.

The family Flaviviridae includes viruses that have different virion structures and morphogenesis mechanisms. Most cellular molecular studies been so far performed with of the Hepacivirus Flavivirus genera. Here, we studied bovine viral diarrhea virus (BVDV), a member Pestivirus genus. We set up method to purify BVDV virions analyzed their morphology by electron microscopy protein lipid composition mass spectrometry. Cryo-electron showed near spherical particles displaying an electron-dense...

We describe a method that can be used to produce equimolar amounts of two or more specific proteins in cell. In this approach, termed the ubiquitin/protein/reference (UPR) technique, reference protein and interest are synthesized as polyprotein separated by ubiquitin moiety. This tripartite fusion is cleaved, cotranslationally nearly so, ubiquitin-specific processing proteases after last residue ubiquitin, producing bearing C-terminal applications such pulse-chase analysis, UPR technique...

Recombinant pestivirus envelope glycoprotein E rns has been shown to interfere with dsRNA-induced interferon (IFN- α / β ) synthesis. This study demonstrated that authentic, enzymically active produced in mammalian cells prevented a IFN response when present the supernatant of bovine cells. Strikingly, synthesis expressing was eliminated after extracellular addition, but not transfection, dsRNA. Importantly, same applied infected viral diarrhea virus (BVDV) lacking N-terminal protease N pro...

ABSTRACT Pestiviruses are the only members of Flaviviridae that encode a nonstructural protease at N terminus their polyproteins. This N-terminal (N pro ) cleaves itself off nascent polyprotein autocatalytically and thereby generates adjacent viral capsid protein C. In previous reports, sequence similarities between catalytic residues papain-like cysteine proteases were put forward. To test this hypothesis, substitutions histidine within carried out by site-directed mutagenesis. Translation...

ABSTRACT The route of internalization the pestivirus bovine viral diarrhea virus (BVDV) was studied by using different chemical and biophysical inhibitors endocytosis. Expression dominant-negative mutant Dyn K44A GTPase dynamin in MDBK cells, as well treatment cells with chlorpromazine β-methyl-cyclodextrin inhibited BVDV entry. infection also abolished potassium (K + ) depletion, hyperosmolarity, endosomal acidification. We conclude that likely enters cell clathrin-dependent endocytosis...

The core protein of pestiviruses is released from the polyprotein by viral and cellular proteinases. Here we report on an additional intramembrane proteolytic step that generates C terminus protein. C-terminal processing classical swine fever virus (CSFV) was blocked inhibitor (Z-LL)(2)-ketone, which specific for signal peptide peptidase (SPP). same effect obtained overexpression dominant-negative SPP D(265)A mutant. presence (Z-LL)(2)-ketone reduced viability CSFV almost 100-fold in a...