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We have analyzed the mechanisms used by adenovirus to gain entry into the host cell. Using both virus infection and the ability of adenovirus particles to enhance polylysine/DNA uptake as a measure of virus entry, we have demonstrated that the adenovirus-encoded 23K protease is required for two functions in the infection process. A proteolytic processing of the capsid is required to generate a virus...
The L2 protein of the human papillomaviruses is a minor structural protein and is not necessary for the major L1 protein to assemble into capsids. However L2 protein binds DNA and enhances the efficiency of HPV capsid assembly, suggesting an important role in the production of infectious papillomaviruses. L2 accumulates rapidly in the nucleus after synthesis in the cytoplasm. To identify L2 sequences...
A comparative study was undertaken to characterize the very early events that distinguish attenuated and pathogenic simian immunodeficiency virus (SIV) infections. Three rhesus macaques were inoculated with the attenuated SIVmac 251 Δnef virus, and three others with a virus of intermediate phenotype, SIVmac 239 nef stop. They were compared to four macaques inoculated with the pathogenic SIVmac 251...
We have analysed the intracellular localisation of herpes simplex virus type 1 ribonucleotide reductase during infection of cultured cells by indirect immunofluorescence using polyclonal and monoclonal antibodies specific for the R1 and R2 subunits. Three different viruses were used to infect cells, wild-type strain 17+and two temperature-sensitive mutants,ts1222, which produces R1 only, andts1207,...
We have determined that HSV causes rapid and large increases in cell-cycle-regulated free E2F and S-phase p107/E2F DNA binding activities in asynchronous cultures of C33A cells. Induction occurred by 4 hr postinfection and coincided with the appearance of viral encoded immediate-early and delayed-early proteins, i.e., when viral DNA replication normally commences. No increase in E2F activities occurred...
The nucleotide sequence of the genes 2 and 3 of theDrosophilarhabdovirus sigma was determined from cDNAs to viral genome and poly(A)+mRNAs. Gene 2 comprises 1032 nucleotides and contains a long ORF encoding a molecular weight 35,208 polypeptide present in infected cells and in virions which migrates in SDS–PAGE as a doublet ofMrabout 60 kDa. The distribution of acidic charges as well as the electrophoretic...
We present evidence that the HIV-1 Tat protein and the RNA-dependent cellular protein kinase, PKR, interact with each other bothin vitroandin vivo.Using GST fusion chromatography, we demonstrate that PKR, interacts directly with the HIV-1 Tat protein. The region in Tat sufficient for binding PKR maps within amino acids 20 to 72. Inin vitroassays, the two-exon form of Tat (Tat 86) was phosphorylated...
The expression of the core gene of two different hepatitis C virus (HCV) isolates was analyzed. In the presence of its downstream E1 envelope protein sequence, two major core protein products with molecular masses of 21 kDa (P21) and 19 kDa (P19) and a minor protein product with molecular mass of 16 kDa (P16) were detected. In the absence of its downstream E1 envelope protein sequence, P21 and P19...
Primate lentiviruses such as human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) are phenotypically diverse, and virus isolates vary in cytopathicity, replication rate, and cell tropism. While all virus isolates infect primary peripheral blood lymphocytes, only a subset of strains infect established CD4-expressing T-cell lines. Here, we describe the development and characterization...
The human immunodeficiency virus type 1 (HIV-1) genome contains 20 exons that are alternatively spliced from 16 splice sites to generate more than 40 different mRNAs, including incompletely spliced and unspliced mRNAs. In contrast to avian retroviral RNA, which has acis-acting element ingagthat negatively regulates splicing (NRS), HIV-1 RNA did not have any NRS sequences in thegagorpolgenes detectable...
Normal rabbit lymphocytes can be infected with HIV-1 although infection is much less efficient than in human lymphocytes. When peripheral blood mononuclear cells (PBMC) from rabbits transgenic for human CD4 (HuCD4) were exposed to HIV-1, enhanced infection and a rapid depletion of lymphocytes were observed. Cell death in the infected cultures occurred via apoptosis, but no similar effect was seen...
The fiber and hexon genes of the immunological adenovirus (Ad) variant strains Ad9/Hx and Ad15/Hx, belonging to subgenus D, were sequenced and compared to the corresponding sequences of the prototypes Ad9 and Ad15. It was found that the variants possessed a novel and identical hemagglutinin as they could not be distinguished by hemagglutination-inhibition tests. These serological data were now confirmed...
Rare cells that contain large amounts of bovine papilloma virus (BPV) DNA have been observed in populations of BPV-transformed mouse ID13 cells. The viral DNA molecules in these “jackpot cells” have been thought to have switched from the controlled replication mode typical of latent BPV infection to the uncontrolled “runaway” prelytic replication characteristic of terminal stage infection of bovine...
The gene encoding the C-terminal protease domain (27 kDa) of the nuclear inclusion protein a of turnip mosaic potyvirus C5 was cloned and expressed as a fusion protein with glutathione S-transferase inEscherichia coliXL1-blue. Two forms of the protease (27 and 25 kDa) were purified from the fusion protein by glutathione affinity chromatography and Mono S chromatography and exhibited the specific proteolytic...
Different serotypes and evolutionary variants of human adenoviruses exhibit distinctive patterns of positive and negative autoregulation of the viral E1A gene. An autoregulatory E1A promoter mutation of the adenovirus type 3 (Ad3) E1A gene renders Ad3hr15 incapable of growth in normally permissive cells. The promoter mutation is complementedin transby E1A products of the heterologous helper adenovirus...
Molecularly cloned SIVmac239 is the prototypical SIVmaclymphocyte-tropic virus that replicates productively in lymphocytes but poorly in macrophages. In macaques, the virus causes activation and productive infection of T lymphocytes which invade the central nervous system (CNS) early after infection in the animal. However, infected animals develop immunosuppression and AIDS but rarely overt neurological...
Expression ofgag/polandenvgenes of human immunodeficiency virus requires the viral Rev protein. Mutant Rev proteins, displaying a transdominant phenotype (TDRev), were shown to inhibit Rev function. To investigate the underlying mechanism of this inhibition, the green fluorescent protein (GFP) ofAequorea victoriawas fused to Rev and TDRev, which allowed the study of their trafficking and interactions...
Molluscum contagiosum virus (MCV) infects preadolescent children and sexually active adults, frequently causing a disfiguring cutaneous disease in immunosuppressed HIV-infected individuals. The development of an efficacious treatment regime has been hampered by the failure to replicate the virus in the laboratory. Here we report the first demonstration of MCV replication in an experimental system...
The phosphoprotein P of human respiratory syncytial virus (RSV) was expressed in eukaryotic cells in phosphorylated form. Site-directed mutagenesis of the recombinant protein established Ser232as the major site of phosphorylationin vivo.Phosphorylation of bacterially made P proteinin vitroby purified casein kinase II (CKII) resulted in the phosphorylation of Ser237, whereas mainly Ser232was phosphorylated...
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