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Epithelial cells are important target cells for coronavirus infection. Earlier we have shown that transmissible gastroenteritis coronavirus (TGEV) and mouse hepatitis coronavirus (MHV) are released from different sides of porcine and murine epithelial cells, respectively. To study the release of these viruses from the same cells, we constructed a porcine LLC-PK1 cell line stably expressing the recombinant...
Previous studies have shown that expression of HIV-1 provirus was enhanced in cells co-infected with the herpes virus and that HSV-1-mediated induction of the HIV-1 provirus expression involved both NF-κB-dependent and NF-κB-independent mechanisms. Nuclear NF-κB complexes could be detected approximately 8 hr after HSV-1 infection. Using NF-κB-specific antibodies in a mobility shift assay, we have...
This study demonstrates that gene-gun inoculation of mice with DNA encoding the influenza virus hemagglutinin (HA) results in the life-long maintenance of protective B cell responses. Using a sensitive single-cell enzyme-linked immunospot assay, we show that all of the HA-specific plasma cells are localized in the bone marrow and spleen 1 year postimmunization. As a consequence of prior virus challenge,...
Retroviruses are expressed under the control of viral control regions designated long terminal repeats (LTRs), which contain all signals for transcriptional initiation as well as transcriptional termination. However, retroviral LTRs from different species within a common genus, such as Lentivirus, do not show significant overall sequence homology. We compiled a model of the functional organization...
The human T-cell leukemia virus type I (HTLV-I) Tax protein and the hepatitis B virus (HBV) X protein have each been shown to activate transcription of their respective viral promoters as well as a subset of cellular gene promoters. Here we show that the HTLV-I long terminal repeat (LTR) is responsive to HBV X transactivation. Maximum levels of X-mediated transactivation of the LTR were 8-fold. An...
During replication of the minute virus of mice (MVM) genome, a dimer replicative form (RF) intermediate is resolved into two monomer RF molecules in such a way as to retain a unique sequence within the left hand hairpin terminus of the viral genome. Although the proposed mechanism for resolution of the dimer RF remains uncertain, it likely involves site-specific nicking of the dimer bridge. The RF...
The aspartyl proteinase (PR) encoded by the feline immunodeficiency virus (FIV) was prepared by total chemical synthesis. The 116-amino-acid polypeptide chain was assembled in a stepwise fashion using a Boc chemistry solid-phase peptide synthesis approach and subsequently folded into the biologically active dimeric proteinase. The synthetic enzyme showed proteolytic activity against a variety of different...
We have previously shown that adenovirus E1A proteins can block interferon-α (IFN-α)-signaling. In the current study, we examined if the same is true for IFN-γ signaling. Cotransfection experiments showed that both 289R and 243R forms of E1A could block the expression of an IFN-γ-inducible reporter gene. Similarly, in an E1A-expressing HeLa cell line IFN-γ failed to induce the synthesis of IRF-1 mRNA...
The expression kinetics of an essentialtrans-regulatory protein, ICP27, from herpes simplex virus type 1 correspond to that of an immediate-early gene whose expression increases rapidly upon infection and then decreases as of 7 hr postinfection. In contrast, here we report that the bovine herpesvirus type 1 (BHV-1) homolog BICP27, a 50-kDa protein, is expressed as an early gene. Both the transcript...
The C1 gene of tomato yellow leaf curl geminivirus (TYLCV) encodes a multifunctional protein (Rep) involved in replication. A truncated form of this gene, capable of expressing the N-terminal 210 amino acids (aa) of the Rep protein, was cloned under the control of the CaMV 35S promoter and introduced intoNicotiana benthamianausingAgrobacterium tumefaciens.The same sequence was also cloned in antisense...
The coding regions for cowpea mosaic virus (CPMV) capsid proteins VP37 and VP23 were introduced separately into a transient plant expression vector containing an enhanced CaMV 35S promoter. Significant expression of either capsid protein was observed only in protoplasts transfected simultaneously with both constructs. Immunosorbent electron microscopy revealed the presence of virus-like particles...
Human MxA protein is a type I interferon-inducible intracytoplasmic protein, which mediates antiviral actions against a variety of negative-strand RNA viruses including influenza A, measles, and vesicular stomatitis viruses. Recently, it has also been shown that several members of theBunyaviridaefamily are inhibited by MxA protein. The hantavirus genus in theBunyaviridaefamily includes important human...
The poliovirus genome comprises a single open reading frame which is translated to give one large polyprotein. The proteolytic cascade involved in the processing of this polyprotein is not yet understood in full detail, particularly concerning the processing of P2–P3, the precursor to the viral nonstructural polypeptides, 2A, 2B, 2C, 3A, 3B, 3C, and 3D. To investigate the possibility that the cleavage...
Replication ofvif-caprine arthritis encephalitis virus (CAEV) is highly attenuated in primary goat synovial membrane cells and blood-derived macrophages compared to the wild-type (wt) virus. We investigated the requirement for CAEV Vif forin vivoreplication and pathogenicity in goats by intra-articular injection of either infectious proviral DNA or viral supernatants. Wild-type CAEV DNA or virus inoculation...
African swine fever virus is an icosahedral double-stranded DNA virus which replicates in the cytoplasm of porcine monocytic cells. Progeny virus particles, like poxviruses and iridoviruses, are assembled in cytoplasmic inclusions, termed virus factories. The formation of these structures in susceptible cells infectedin vitrowith pathogenic and attenuated ASFV isolates has been studied by semiquantitative...
Homologous regions (hrs), which are present at eight dispersed locations on theAutographa californicamultinucleocapsid nuclear polyhedrosis virus genome, are composed of repeated imperfect palindromes within directly repeated sequences.Hrsact as transcriptional enhancers of RNA polymerase II-mediated transcription and as origins of DNA replication when incorporated into plasmids and tested in transient...
We compared the amino acid sequences of groups of receptor (CD46) downregulating and nondownregulating measles virus (MV) hemagglutinins (Hs) and identified seven group-specific differences as candidates for the mediation of the observed differential effects. Using site-directed mutagenesis, we mutated the chosen amino acids of the H of MV-strain WTF (WTF-H), a nondownregulating H, and introduced...
Identification of regulators of p53 expression is a crucial step in understanding the diverse functions of p53 and its role in cellular homeostasis and responsiveness to insult. Several viral proteins inactivate p53 as a modulator of cell cycle progression and apoptosis. Here, we report that a unique leporipoxviral transcription factor greatly increases levels of p53 mRNA. C7, an early transcription...
MuLV-induced myeloid leukemias (MML) having promonocytic characteristics are produced with high incidence in some strains of adult mice that are undergoing chronic peritoneal inflammation. Previously we showed that many leukemias have rearrangements of the c-myblocus due to insertional mutagenesis, however, we also identified a number of leukemias that had proviral integrations in the absence of c-mybrearrangement...
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