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Journal of Clinical Microbiology Apr 1978Simian papovavirus SA12 agglutinated human, guinea pig, and chicken erythrocytes. SA12 hemagglutinin was most effectively released from debris of infected tissue culture...
Simian papovavirus SA12 agglutinated human, guinea pig, and chicken erythrocytes. SA12 hemagglutinin was most effectively released from debris of infected tissue culture cells at an alkaline pH.
Topics: Animals; Antibody Formation; BK Virus; Cross Reactions; Haplorhini; Hemagglutination, Viral; Hydrogen-Ion Concentration; Polyomavirus; Simian virus 40
PubMed: 29051
DOI: 10.1128/jcm.7.4.396-398.1978 -
Uirusu Jun 2002
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Infection and Immunity Oct 1980Paired sera from 150 pregnant women and 387 umbilical cord sera were tested for BK virus (BKV) antibodies. The hemagglutination inhibition, neutralization, and indirect...
Paired sera from 150 pregnant women and 387 umbilical cord sera were tested for BK virus (BKV) antibodies. The hemagglutination inhibition, neutralization, and indirect immunofluorescence tests were employed for the detection of antibodies. Treatment of serum with anti-gamma Fc and tests of immunoglobulin M (IgM) fractions for antibodies were utilized as required to detect and validate the presence of virus-specific IgM. The BKV antibody prevalence in the sera collected at the time of the first prenatal visit was 75% by hemagglutination inhibition and 91% by neutralization tests. A total of 95% of the women had antibodies by at least one of the three serological tests. Five of 100 women with normal pregnancies exhibited BKV activity during pregnancy as evidenced by a greater than fourfold rise in BKV hemagglutination inhibition antibody titers and acquisition of BKV-specific IgM. The antibody rise occurred in the younger women and appeared to be a result of reactivation of the virus rather than of primary infection. Two instances of possible recent BKV infections were identified. BKV-specific IgM was not detected in any of the 387 umbilical cord sera which included three specimens from infants born to mothers with definite or probable BKV activity during pregnancy and 50 specimens with IgM levels of > 20 mg/100 ml. The results indicate that few women in the child-bearing age are nonimmune to BKV and that, although reactivation of infection occurs in pregnancy, congenital transmission of the virus either does not occur or is rare.
Topics: Animals; Antibodies, Viral; BK Virus; Female; Fetal Blood; Fluorescent Antibody Technique; Hemagglutination Inhibition Tests; Humans; Immunoglobulin M; Neutralization Tests; Polyomavirus; Pregnancy; Pregnancy Complications, Infectious; Tumor Virus Infections
PubMed: 6254883
DOI: 10.1128/iai.30.1.29-35.1980 -
Journal of Virology May 2000Small DNA tumor viruses like human papillomaviruses, simian virus 40, and adenoviruses modulate the activity of cellular tumor suppressor proteins p53 and/or pRB. These...
Small DNA tumor viruses like human papillomaviruses, simian virus 40, and adenoviruses modulate the activity of cellular tumor suppressor proteins p53 and/or pRB. These viruses replicate as nuclear multicopy extrachromosomal elements during the S phase of the cell cycle, and it has been suggested that inactivation of p53 and pRb is necessary for directing the cells to the S phase. Mouse polyomavirus (Py), however, modulates only the pRB protein activity without any obvious interference with the action of p53. We show here that Py replication was not suppressed by the p53 protein indeed in all tested different mouse cell lines. In addition, E1- and E2-dependent papillomavirus origin replication was insensitive to the action of p53 in mouse cells. We show that in hamster (Chinese hamster ovary) or human (osteosarcoma 143) cell lines the replication of both Py and papillomavirus origins was efficiently blocked by p53. The block of Py replication in human and hamster cells is not caused by the downregulation of large T-antigen expression. The deletion analysis of the p53 protein shows that the RPA binding, proline-rich regulatory, DNA-binding, and oligomerization domains are necessary for p53 action in both replication systems. These results indicate that in mouse cells the p53 protein could be inactive for the suppression of papovavirus replication.
Topics: Animals; Bovine papillomavirus 1; CHO Cells; Cell Line; Cricetinae; Fibroblasts; Gene Expression Regulation, Viral; Humans; Immunoblotting; Mice; Plasmids; Polyomavirus; Polyomavirus Infections; Replication Origin; Transfection; Tumor Cells, Cultured; Tumor Suppressor Protein p53; Tumor Virus Infections; Virus Replication
PubMed: 10775606
DOI: 10.1128/jvi.74.10.4688-4697.2000 -
Journal of Virology Sep 1988BK virus (BKV) is a human papovavirus which latently infects a majority of the world population. Reactivation of this virus is associated with acute hemorrhagic...
BK virus (BKV) is a human papovavirus which latently infects a majority of the world population. Reactivation of this virus is associated with acute hemorrhagic cystitis, and BKV DNA has been found in human tumor tissue. BKV is one of many highly homologous papovaviruses, including simian virus 40 and JC virus, which display distinct host and cell-type specificities, transformation potentials, and pathologies. These differences are thought to be determined, in part, by the noncoding regulatory region of each virus, which contains the origin of replication and regulatory elements for both early and late gene expression. We have used linker scan mutants to map functional elements of a truncated BKV early promoter and enhancer and have studied the stereospecific requirements of these elements. We have also identified protein-binding regions through DNase protection studies. Our results show that a minimum of four elements are necessary for efficient early transcription, at least three of which correspond to DNase-protected domains. These protein-binding elements map to the TATA box and two nuclear factor 1 consensus sequences, one located within the enhancer repeat unit and the other located to the late side of the enhancer. The sequence of the fourth element is similar to the transcription factor Sp1 consensus sequence. Additional DNase-protected sites are centered over AP-1 and Sp1 consensus sequences. Finally, we find that the functional elements of the BKV early promoter and enhancer lack strict stereospecific requirements for efficient transcription.
Topics: BK Virus; Base Sequence; DNA, Viral; Enhancer Elements, Genetic; HeLa Cells; Humans; Molecular Sequence Data; Mutation; Plasmids; Polyomavirus; Promoter Regions, Genetic; Regulatory Sequences, Nucleic Acid; Repetitive Sequences, Nucleic Acid; Transcription, Genetic
PubMed: 2841491
DOI: 10.1128/JVI.62.9.3378-3387.1988 -
Journal of Virology Aug 1980Human papovavirus JC virus was adapted to growth in human embryonic kidney (HEK) cells. After eight passages, the HEK-adapted JC virus produced high virus yields and was...
Human papovavirus JC virus was adapted to growth in human embryonic kidney (HEK) cells. After eight passages, the HEK-adapted JC virus produced high virus yields and was capable of forming plaques in HEK monolayer cultures. Eleven plaque-purified stocks were prepared and characterized. Biologically, the plaque-purified virus induced tumor and viral antigens in HEK cells earlier and in a higher percentage of cells than uncloned virus. Cytopathic changes were also evident sooner and were more extensive. The DNA from uncloned as well as plaque-purified isolates was analyzed by restriction endonuclease cleavage followed by gel electrophoresis. The DNA from uncloned HEK-adapted virus was heterogeneous. Plaque-purified virus isolates yielded DNA which, although much less heterogeneous than the uncloned stock, still consisted of two or more species of viral DNA.
Topics: Cells, Cultured; DNA Restriction Enzymes; DNA, Viral; Embryo, Mammalian; Humans; Kidney; Polyomavirus; Viral Plaque Assay; Virus Cultivation; Virus Replication
PubMed: 6255188
DOI: 10.1128/JVI.35.2.498-504.1980 -
Virology Jan 2016Polyomaviruses induce cell proliferation and transformation through different oncoproteins encoded within the early region (ER): large T antigen (LT), small T antigen...
Polyomaviruses induce cell proliferation and transformation through different oncoproteins encoded within the early region (ER): large T antigen (LT), small T antigen (sT) and, in some cases, additional components. Each virus utilizes different mechanisms to achieve transformation. For instance, the LTs of Simian virus 40 (SV40), BK and/or JC virus can induce transformation; but Merkel Cell Polyomavirus (MCPyV) requires expression of sT. Lymphotropic Papovavirus (LPV) is closely related to Human Polyomavirus 9 (HuPyV9) and, under similar conditions, mice expressing LPV.ER exhibit higher rates of tumor formation than mice expressing SV40.ER. We have investigated the contributions of individual LPV.ER components to cell transformation. In contrast to SV40, LPV.ER transforms mouse embryonic fibroblasts (MEFs), but expression of LPV LT is insufficient to transform MEFs. Furthermore, LPV sT induces immortalization and transformation of MEFs. Thus, in the case of LPV, sT is the main mediator of oncogenesis.
Topics: Animals; Antigens, Viral, Tumor; Cell Transformation, Neoplastic; Cells, Cultured; Fibroblasts; Mice; Polyomavirus; Retinoblastoma-Like Protein p107; Retinoblastoma-Like Protein p130; Tumor Suppressor Protein p53
PubMed: 26517398
DOI: 10.1016/j.virol.2015.10.003 -
The EMBO Journal Jan 1992Simian virus 40 (SV40) and polyomavirus (Py) DNA replication require cellular proteins and a virus-encoded early gene product, large T antigen (SVT and PyT,... (Comparative Study)
Comparative Study
Simian virus 40 (SV40) and polyomavirus (Py) DNA replication require cellular proteins and a virus-encoded early gene product, large T antigen (SVT and PyT, respectively). Primate cells contain factors permissive for SV40 replication, whereas murine cells express those factors permissive for Py. We have compared the roles T antigen, cell permissiveness and replication play in transcription of SV40 and Py genes. We show that in their respectively permissive cells, SV40 replication causes a major shift in transcription initiation from the early to the late viral promoter, whereas when Py replicates a comparable shift does not occur. This difference is discussed in relation to differences in the organization of the origin and promoter region between these two papovaviruses. Reporter plasmids were constructed that carried both viral origins, one at the natural position in the promoter being tested and the other at a distal location. With the appropriate TAg, these vectors could be made to replicate in either primate (HeLa) or rodent (3T6) cells. The SV40 early to late shift occurred when replication was driven in HeLa cells, and was not seen on replicating templates in rodent cells. Thus, replication per se does not account for the shift. We show also that, like SVT, PyT is a potent activator of transcription, and that SVT and PyT can activate each other's late promoters independently of DNA replication, but only in cells permissive for DNA replication catalysed by the respective T antigen. Taken together, the data presented here suggest that papovaviruses may utilize permissive factors in transcription control mechanisms.
Topics: Animals; Antigens, Viral, Tumor; Cell Line; DNA Replication; Gene Expression Regulation, Viral; Humans; Mice; Papillomaviridae; Polyomaviridae; Polyomavirus; Promoter Regions, Genetic; Simian virus 40; Transcription, Genetic; Transcriptional Activation
PubMed: 1310931
DOI: 10.1002/j.1460-2075.1992.tb05040.x -
Journal of Virology Sep 1988The human papovavirus BK has a noncoding regulatory region located between the divergently transcribed early and late coding regions. Many strains of BK virus (BKV) have...
The human papovavirus BK has a noncoding regulatory region located between the divergently transcribed early and late coding regions. Many strains of BK virus (BKV) have direct DNA sequence repeats in the regulatory region, although the number and extent of these repeats varies widely between independent isolates. Until recently, little was known about the individual functional elements within the BKV regulatory region, and the biological significance of the variable repeat structure has been unclear. To characterize the interaction between sequences in the BKV regulatory region and host cell transcription factors, we have carried out DNase I footprinting and competitive binding experiments on three strains of BKV, including one strain that does not contain direct sequence repeats. We have used relatively crude fractions from HeLa cell nuclear extracts, as well as DNA affinity-purified preparations of proteins. Our results demonstrate that BK(Dunlop), BK(WW), and BK(MM) each contain multiple binding sites for a factor, NF-BK, that is a member of the nuclear factor 1 family of transcription factors. We predict the presence of three to eight binding sites for NF-BK in the other strains of BKV for which a DNA sequence is available. This suggests that the binding of this protein is likely to be required for biological activity of the virus. In addition to NF-BK sites, BK(WW) and BK(MM) each contain a single binding site for transcription factor Sp1, and BK(Dunlop) contains two binding sites for transcription factor AP-1. The AP-1 sites in BK(Dunlop) span the junction of adjacent direct repeats, suggesting that repeat formation may be an important mechanism for de novo formation of binding sites not present in a parental strain.
Topics: BK Virus; Binding, Competitive; CCAAT-Enhancer-Binding Proteins; Chromatography, Affinity; DNA, Viral; DNA-Binding Proteins; Deoxyribonuclease I; HeLa Cells; Humans; NFI Transcription Factors; Nuclear Proteins; Polyomavirus; Regulatory Sequences, Nucleic Acid; Repetitive Sequences, Nucleic Acid; Transcription Factors; Y-Box-Binding Protein 1
PubMed: 2841492
DOI: 10.1128/JVI.62.9.3388-3398.1988 -
Journal of Virology Nov 1995Binding of B-lymphotropic papovavirus (LPV) to host cells differing in susceptibility to viral infection was determined by a newly established, direct, nonradioactive...
Binding of B-lymphotropic papovavirus (LPV) to host cells differing in susceptibility to viral infection was determined by a newly established, direct, nonradioactive virus binding assay, which allows quantitative description of the binding characteristics by receptor saturation and Scatchard analysis. LPV binding to the highly susceptible human B-lymphoma cell line BJA-B K88 is specific, saturable, and noncooperative. Binding occurs very fast, with an association rate constant (k1) of 6.7 x 10(7) M-1s-1, and is of high affinity, with a dissociation constant (Kd) of 2.9 x 10(-12) M; and the virus-receptor complex is stable, with a half life of 70 min. The binding affinities of receptors on four other highly, moderately, or weakly susceptible human B-lymphoma cell lines were similar, with up to twofold variation around a mean Kd value of 3 x 10(-12) M, suggesting the presence of the same LPV receptor on all of these cell lines. This view is further supported by the finding that in all cases a terminal sialic acid is necessary for LPV binding. Tunicamycin has been shown to drastically induce LPV susceptibility and LPV binding in weakly and moderately susceptible B-lymphoma cell lines (O.T. Keppler, M. Herrmann, M. Oppenländer, W. Meschede, and M. Pawlita, J. Virol. 68:6933-6939, 1994). The hypothesis that the constitutively expressed and tunicamycin-induced LPV receptors are identical is strengthened by our finding that both receptor types displayed the same high affinity. LPV susceptibility of different B-lymphoma cell lines was correlated with receptor number but not with receptor affinity. The numbers of receptors per cell on highly and moderately susceptible cell lines ranged from 2,000 to 400 and were directly proportional to LPV susceptibility. This indicates that the number of high-affinity receptors per cell is a key regulating factor for the LPV host range.
Topics: Animals; B-Lymphocytes; Cell Line; Cricetinae; Enzyme-Linked Immunosorbent Assay; Humans; Kinetics; Lymphoma, B-Cell; Mathematics; Mice; Models, Theoretical; Neuraminidase; Papillomaviridae; Polyomaviridae; Rats; Receptors, Virus; Species Specificity; Tumor Cells, Cultured; Tunicamycin
PubMed: 7474091
DOI: 10.1128/JVI.69.11.6797-6804.1995