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1.
Science ; 196(4286): 183-5, 1977 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-191907

RESUMO

The detection and recovery of SV40 genomes containing foreign DNA sequences can be facilitated, and the risk of accidental dispersal reduced, by in situ hybridization and radioautography.


Assuntos
DNA Recombinante/análise , DNA Viral/análise , Hibridização de Ácido Nucleico , Vírus 40 dos Símios/análise , Linhagem Celular , Colífagos/análise
2.
Microbiol Mol Biol Rev ; 65(2): 288-318 ; second and third pages, table of contents, 2001 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11381103

RESUMO

"It has been commented by someone that 'polyoma' is an adjective composed of a prefix and suffix, with no root between--a meatless linguistic sandwich" (C. J. Dawe). The very name "polyomavirus" is a vague mantel: a name given before our understanding of these viral agents was clear but implying a clear tumor life-style, as noted by the late C. J. Dawe. However, polyomavirus are not by nature tumor-inducing agents. Since it is the purpose of this review to consider the natural function of middle T antigen (MT), encoded by one of the seemingly crucial transforming genes of polyomavirus, we will reconsider and redefine the virus and its MT gene in the context of its natural biology and function. This review was motivated by our recent in vivo analysis of MT function. Using intranasal inoculation of adult SCID mice, we have shown that polyomavirus can replicate with an MT lacking all functions associated with transformation to similar levels to wild-type virus. These observations, along with an almost indistinguishable replication of all MT mutants with respect to wild-type viruses in adult competent mice, illustrate that MT can have a play subtle role in acute replication and persistence. The most notable effect of MT mutants was in infections of newborns, indicating that polyomavirus may be highly adapted to replication in newborn lungs. It is from this context that our current understanding of this well-studied virus and gene is presented.


Assuntos
Antígenos Transformantes de Poliomavirus/fisiologia , Polyomavirus/fisiologia , Animais , Apoptose , Diferenciação Celular , Transformação Celular Viral , Células Eucarióticas/virologia , Camundongos , Camundongos Knockout , Polyomavirus/imunologia , Transdução de Sinais , Transcrição Gênica , Replicação Viral
3.
Curr Top Microbiol Immunol ; 299: 349-70, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16568906

RESUMO

The Parvoviridae, a family of viruses with single-stranded DNA genomes widely spread from invertebrates to mammal and human hosts, display a remarkable evolutionary capacity uncommon in DNA genomes. Parvovirus populations show high genetic heterogeneity and large population sizes resembling the quasispecies found in RNA viruses. These viruses multiply in proliferating cells, causing acute, persistent or latent infections relying in the immunocompetence and developmental stage of the hosts. Some parvovirus populations in natural settings, such as carnivore autonomous parvoviruses or primate adeno associated virus, show a high degree of genetic heterogeneity. However, other parvoviruses such as the pathogenic B19 human erythrovirus or the porcine parvovirus, show little genetic variation, indicating different virus-host relationships. The Parvoviridae evolutionary potential in mammal infections has been modeled in the experimental system formed by the immunodeficient scid mouse infected by the minute virus of mice (MVM) under distinct immune and adaptive pressures. The sequence of viral genomes (close to 10(5) nucleotides) in emerging MVM pathogenic populations present in the organs of 26 mice showed consensus sequences not representing the complex distribution of viral clones and a high genetic heterogeneity (average mutation frequency 8.3 x 10(-4) substitutions/nt accumulated over 2-3 months). Specific amino acid changes, selected at a rate up to 1% in the capsid and in the NS2 nonstructural protein, endowed these viruses with new tropism and increased fitness. Further molecular analysis supported the notion that, in addition to immune pressures, the affinity of molecular interactions with cellular targets, as the Crml nuclear export receptor or the primary capsid receptor, as well as the adaptation to tissues enriched in proliferating cells, are major selective factors in the rapid parvovirus evolutionary dynamics.


Assuntos
Infecções por Parvoviridae/virologia , Parvovirus/genética , Vírus de RNA/genética , Animais , Capsídeo/química , Evolução Molecular , Variação Genética , Genoma Viral , Humanos , Parvovirus/imunologia , Replicação Viral
4.
Mol Cell Biol ; 6(6): 2068-79, 1986 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-3023917

RESUMO

Heterologous enhancer recombinants and deletions of the polyomavirus (Py) noncoding region were constructed and analyzed for tissue specificity of DNA replication and transcription in a number of lymphoid and other cell lines. The simian virus 40 72-base-pair repeat, mouse immunoglobulin heavy-chain enhancer, and Moloney murine leukemia virus enhancer were inserted into the PvuII-D locus (nucleotides 5128 through 5265) of Py. The ability of these recombinants and the parental PvuII-D deletion mutant to replicate in permissive 3T6 cells and MOP-6 cells as well as in nonpermissive mouse B lymphoid, T lymphoid, mastocyte, and embryonal carcinoma cells was determined. Wild-type Py DNA was not permissive for replication in most lymphoid cell lines, except one hybridoma line. Simply deleting the Py PvuII-D region, however, gave Py an expanded host range, allowing high-level replication in some T lymphoid and mastocytoma cell lines, indicating that this element can be a tissue-specific negative as well as positive element. Substitution of the murine leukemia virus enhancer for Py PvuII-D yielded a Py genome which retained the ability to replicate in 3T6 cells but also replicated well in B lymphoid cells. Substitution with the immunoglobulin heavy-chain enhancer allowed replication in B lymphoid cells but interfered with replication in 3T6 cells and mastocytomas. Surprisingly, substitution with the simian virus 40 72-base-pair enhancer repeat gave a recombinant which would not replicate in any cell line tried, including MOP-6 cells, even though other recombinants with this enhancer would replicate. Thus, we observed both cooperation and interference in these combinations between enhancer components and the Py genome and that these combined activities were cell specific. These results are presented as evidence that there may be a positional dependence, or syntax, for the recognition of genetic elements controlling Py tissue specificity.


Assuntos
Replicação do DNA , Elementos Facilitadores Genéticos , Genes Reguladores , Polyomavirus/genética , Transcrição Gênica , Animais , Linhagem Celular , Regulação da Expressão Gênica , Camundongos , Vírus 40 dos Símios/genética , Distribuição Tecidual , Replicação Viral
5.
Mol Cell Biol ; 8(5): 1993-2004, 1988 May.
Artigo em Inglês | MEDLINE | ID: mdl-2838739

RESUMO

Polyomavirus (Py) enhancer core elements were compared for their ability to activate Py early transcription and DNA replication in mouse 3T6 cells, lymphoid cell lines, and undifferentiated embryonal carcinoma cells. By examining the pattern of genetic change in a number of cell-specific Py variants, we identified subenhancer sequences that may be functionally important for virus replication. Four such distinct enhancer consensus sequences were synthesized and designated as the A core (homologous with adenovirus 5 E1A enhancer), B core (homologous to the simian virus 40 A enhancer core), C core (containing an inverted repeat within the Py B enhancer), and BPV core (homologous to the bovine papillomavirus enhancer). When used to replace the complete Py B enhancer, single copies of all but the BPV element were able to fully activate Py DNA replication after transfection, but this activation was usually cell type specific. In the PCC4 embryonal carcinoma cells, only the A-core sequence was able to activate transcription and DNA replication. The BPV core sequence containing the Py F441 point change was unable to activate DNA replication in the F9 embryonal carcinoma or any other cell line. No single insertion element was dominant nor did these elements display the wild-type enhancer pattern of cell-specific activation of DNA replication. In addition, differential effects were often observed on the activation of transcription versus DNA replication. In 3T6 cells, transcription could be highly activated by the A core without a corresponding activation of DNA replication. In murine T lymphoid cell lines, the B core activated DNA replication without a corresponding increase in transcription. Furthermore, both DNA replication and, to a lesser degree, transcription often showed a strong tissue-specific dependence on the polarity of the inserted core element for activation.


Assuntos
Replicação do DNA , Elementos Facilitadores Genéticos , Genes Virais , Polyomavirus/genética , Transcrição Gênica , Replicação Viral , Animais , Sequência de Bases , Linhagem Celular , Fibroblastos/metabolismo , Leucócitos/metabolismo , Camundongos , Dados de Sequência Molecular , Especificidade de Órgãos , Polyomavirus/fisiologia , Teratoma/patologia , Células Tumorais Cultivadas/metabolismo
6.
Mol Cell Biol ; 3(8): 1381-8, 1983 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-6312290

RESUMO

A late region deletion mutant of simian virus 40 (dl5) was previously shown to be deficient in the transport of nuclear RNA. This is a splice junction deletion that has lost the 3' end of an RNA leader, an intervening sequence, and the 5' end of the splice acceptor site on the body of the mRNA. In this report, we analyzed the steady-state structure of the untransported nuclear RNA. The 5' ends of this RNA are heterogeneous but contain a prominent 5' end at the normal position (nucleotide 325) in addition to several other prominent 5' ends not seen in wild-type RNA. The 3' end of this RNA does not occur at the usual position (nucleotide 2674) of polyadenylation; instead, this RNA is non-polyadenylated, with the 3' end occurring either downstream or upstream of the normal position.


Assuntos
Núcleo Celular/metabolismo , Poli A/genética , Splicing de RNA , Animais , Transporte Biológico , Compartimento Celular , Células Cultivadas , Deleção Cromossômica , Haplorrinos , Mutação , RNA Viral/genética , RNA Viral/metabolismo , Vírus 40 dos Símios/genética
7.
Mol Cell Biol ; 2(12): 1550-7, 1982 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-14582196

RESUMO

The construction of a recombinant virus in the late region of simian virus 40 is presented. The small intervening sequence of late 19S RNA (0.760 to 0.765 map unit) was cloned and inserted into the EcoRI site (1.0 map unit) in the late region of simian virus 40. This is a mutant virus that now has two intervening sequences, one at the normal position (0.760 map unit) and another out of the context of its flanking sequence and now at 1.0 map unit. The recombinant appears poisonous, as repeated attempts to plaque it as a virus with a standard helper virus were unsuccessful. The transcription of this recombinant was, therefore, studied after direct DNA transfection onto CV-1 cells. Nuclease S1 analysis of mutant RNA indicates that the major nuclear transcript was a spliced but nuclear 16S RNA species. Normally, 16S RNA is not found in the nucleus. This result was shown to be an artifact of the DNA transfection protocol. When the glycerol shock was done after infection with virus, a similar alteration in the makeup of nuclear RNA was seen. A transient stock of this double-intron mutant was finally obtained, using a nonrevertable helper virus. The transcriptional analysis of this mutant showed that unspliced 19S RNA was not transported and remained within the nucleus, whereas spliced 19S and 16S RNAs were transported. We conclude that the retention of nuclear transcripts within the nucleus is not simply due to the presence of intronic sequences, as spliced 19S and 16S RNAs which contain the second intron were efficiently transported.


Assuntos
Íntrons , RNA Nuclear/metabolismo , Vírus 40 dos Símios , Transporte Ativo do Núcleo Celular , RNA Nuclear/genética , RNA Viral/metabolismo , Vírus 40 dos Símios/genética , Vírus 40 dos Símios/metabolismo
8.
Mol Cell Biol ; 5(6): 1534-7, 1985 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-2993870

RESUMO

The simian virus 40 72-base-pair repeats substituted for the polyomavirus enhancer, allowing replication and transcription in mouse 3T6 but not monkey CV-1 cells. A polyomavirus genome containing the entire simian virus 40 control region replicated at low levels in 3T6 and CV-1 cells; however, transcripts were detected only in 3T6 cells. Our results suggest that the simian virus 40 72-base-pair repeats are unable to alter the host species specificity of the complete polyomavirus genome.


Assuntos
Replicação do DNA , Elementos Facilitadores Genéticos , Genes Reguladores , Polyomavirus/genética , Vírus 40 dos Símios/genética , Replicação Viral , Animais , Linhagem Celular , Chlorocebus aethiops , Genes Virais , Camundongos , Sequências Repetitivas de Ácido Nucleico , Especificidade da Espécie , Transcrição Gênica
9.
Mol Cell Biol ; 10(9): 4996-5001, 1990 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-2167444

RESUMO

The cell-specific regulation of DNA replication has important implications for the molecular strategy of cellular gene control. Mouse polyomavirus (Py) DNA replication is examined as a model of cell-specific replication control. Using an FM3A-derived mouse cell line which expresses early viral proteins (FOP cells), we determined the minimal sequence requirements for viral DNA replication. FOP cells were observed to have much simpler enhancer requirements than 3T6 and many other cells and did not need a B enhancer for high levels of DNA replication. Using these cells, we show that the individual or tandem binding sites for several unrelated trans-acting factors which are generally subfunctional as transcriptional enhancers (simian virus 40 A core, TGTGGAATG; EBP20, TGTGGTTTT; PEA1 [an AP-1 analog], GTGACTAA; PEA2, GACCGCAG; and PEA3, AGGAAG) stimulated low levels of Py DNA replication. The ordered dimeric combination of PEA3 and PEA1 factor-binding sites, however, acted synergistically to stimulate viral DNA replication to high wild-type levels. This is in contrast to prior results in which much larger enhancer sequences were necessary for high-level viral DNA replication. PEA3/PEA1-stimulated DNA replication showed a distance and orientation independence relative to the origin, which disagrees with some but not other prior analyses of enhancer-dependent DNA replication. It therefore appears that trans-acting factor-binding sites (enhansons) can generally activate DNA replication and that the AP-1 family of sites may act synergistically with other associated trans-acting factors to strongly affect Py DNA replication in specific cells.


Assuntos
Replicação do DNA , Proteínas de Ligação a DNA/metabolismo , Elementos Facilitadores Genéticos , Polyomavirus/genética , Vírus 40 dos Símios/genética , Fatores de Transcrição/metabolismo , Animais , Sequência de Bases , Sítios de Ligação , Linhagem Celular , Subunidade alfa 1 de Fator de Ligação ao Core , Vetores Genéticos , Camundongos , Dados de Sequência Molecular , Proteínas Tirosina Quinases/metabolismo , Proteínas Proto-Oncogênicas c-jun , Fator de Transcrição AP-2
10.
Mol Cell Biol ; 5(12): 3644-6, 1985 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-3870134

RESUMO

Expression of the gene encoding transforming growth factor-alpha (TGF alpha) was examined in developing rat embryos by using a cloned TGF alpha cDNA as a hybridization probe. Northern blot analysis of RNA isolated from whole fetuses revealed that TGF alpha mRNA was present at relatively high levels in 8- through 10-day-old embryos and then declined to the low or undetectable level, which is characteristic of adult tissues before birth. The level of TGF alpha mRNA present during early gestation was similar to that present in retrovirus-transformed cells in culture, suggesting that TGF alpha expression is not highly localized in the embryo. These observations are consistent with the hypothesis that TGF alpha plays a role in development, possibly as a fetal growth factor.


Assuntos
Peptídeos/genética , RNA Mensageiro/genética , Animais , Embrião de Mamíferos/metabolismo , Regulação da Expressão Gênica , Idade Gestacional , Biossíntese Peptídica , Ratos , Fatores de Crescimento Transformadores
11.
J Virol Methods ; 56(2): 173-8, 1996 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-8882647

RESUMO

Py DNA replicative intermediates (RIs) were mapped using a neutral/neutral two-dimensional (N/N 2D) technique by both chemiluminescent and radioactive detection. The nonradioactive method provides similar sensitivity to radioactive methods while possessing the advantages of stability of labeled probes, faster processing, multiple exposures, easy disposal and safety associated with nonradioactive detection. Application of this method to 2D gel analysis requires some technical modification to reduce background and save the substrate. The patterns of Py RIs with Afl II digestion suggest that bidirectional replication occurs during Py DNA synthesis in cell culture, but other unexpected structures of DNA replication are also seen.


Assuntos
DNA Viral/análise , Eletroforese em Gel Bidimensional/métodos , Polyomavirus/genética , Animais , Linhagem Celular , DNA Viral/metabolismo , Digoxigenina/química , Medições Luminescentes , Camundongos , Sensibilidade e Especificidade
12.
Brain Res ; 736(1-2): 297-304, 1996 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-8930336

RESUMO

Replication-deficient adenoviral recombinants were assessed for in vivo transduction of rat hippocampal CA1 cells. Results show that efficient widespread transduction of CA1 in vivo was rapidly achievable and was sustained for more than 5 weeks. Assessment of electrophysiological properties in acute hippocampal slices showed that synaptic functioning and mechanisms involved in long-term potentiation (LTP) were preserved for minimally 5 weeks postinfection. Hence, adenovirus-mediated gene transfer in vivo promises to be a valuable tool for dissecting molecular mechanisms of synaptic plasticity, such as LTP and long-term depression (LTD).


Assuntos
Adenovírus Humanos , Hipocampo/fisiologia , Neurônios/fisiologia , Sinapses/fisiologia , beta-Galactosidase/biossíntese , Animais , Linhagem Celular , Estimulação Elétrica , Genes Reporter , Hipocampo/citologia , Humanos , Técnicas In Vitro , Potenciação de Longa Duração , Masculino , Plasticidade Neuronal , Neurônios/citologia , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes de Fusão/biossíntese , Transfecção
13.
Magn Reson Imaging ; 18(3): 311-7, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10745141

RESUMO

We studied the changes of tumor size after gene therapy treatment and its relationship with the changes of vascular volume as measured by dynamic contrast-enhanced magnetic resonance imaging (MRI), to investigate whether the vascular changes is predictive of tumor regression. The study was carried out using a spontaneously regressing rat tumor model (C6 Glioma grown subcutaneously in rats). Three rats were treated with recombinant adenoviruses expressing three genes, mouse interleukin 1-alpha (IL1-alpha), mouse interferon gamma (IFN-gamma), and human transforming growth factor beta (TGF-beta), one from each kind. Two rats were treated with saline as controls. Longitudinal studies were performed to monitor the changes of tumor volume (based on T(2)-weighted images) and the vascular volume (based on dynamic contrast enhanced images). In untreated animals, tumor regression was preceded by several days with a decrease in vascular volume. When the tumor growth was perturbed by expression of mouse IL-1alpha, the increase in vascular volume was correlated with the continuing growth in size, and the decrease in vascular volume was predictive of the onset of tumor regression. As new advances in immunotherapy in cancer treatment emerge, the ability to determine the efficacy of therapy as early as possible will enable optimization of treatment regiments. The vascularity changes measured by dynamic MRI may provide a means to serve for this purpose.


Assuntos
Terapia Genética , Glioma/patologia , Glioma/terapia , Aumento da Imagem/métodos , Angiografia por Ressonância Magnética/métodos , Neovascularização Patológica/diagnóstico , Adenoviridae/genética , Animais , Gadolínio DTPA/farmacocinética , Vetores Genéticos/genética , Vetores Genéticos/farmacologia , Glioma/irrigação sanguínea , Glioma/metabolismo , Humanos , Interferon gama/biossíntese , Interferon gama/genética , Interferon gama/farmacologia , Interleucina-1/biossíntese , Interleucina-1/genética , Interleucina-1/farmacologia , Fígado/irrigação sanguínea , Fígado/metabolismo , Estudos Longitudinais , Camundongos , Microcirculação/efeitos dos fármacos , Regressão Neoplásica Espontânea , Transplante de Neoplasias , Neovascularização Patológica/metabolismo , Dinâmica não Linear , Ratos , Ratos Wistar , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/farmacologia , Proteínas Recombinantes , Indução de Remissão , Fator de Crescimento Transformador beta/biossíntese , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/farmacologia , Células Tumorais Cultivadas
14.
Med Hypotheses ; 55(1): 1-4, 2000 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11021317

RESUMO

The generation and accumulation of genetic mutations have been associated with carcinogenesis. Epidemiological and experimental evidence implicate parvoviruses in growth inhibition (oncosuppression) and selective destruction (oncolysis) of tumor cells. It is proposed here that parvoviruses can preferentially target genetically unstable tumor cells, which are deficient in DNA repair mechanisms. This selective strategy may serve as a virus-based therapeutic approach against cancer.


Assuntos
Neoplasias/prevenção & controle , Infecções por Parvoviridae/fisiopatologia , Parvovirus/genética , Animais , Humanos , Neoplasias/epidemiologia , Infecções por Parvoviridae/epidemiologia , Células Tumorais Cultivadas
15.
Med Hypotheses ; 47(1): 1-9, 1996 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-8819107

RESUMO

Genetic instability characterized by the accumulation of mutations of tumor suppressor genes and oncogenes appears to be associated with carcinogenesis in colorectal and other cancers. Mutations of DNA polymerase beta (pol beta) and related chromosomal alterations appear to be consistent with the causal role of a "mutator phenotype' in carcinogenesis. However, homozygous knockout pol beta mutations appear to interfere with embryogenesis. Increased pol beta activity (i.e. relative to pol alpha activity) has been associated with cell cycle arrest. The related aphidicolin-resistant DNA replication has been observed primarily in differentiating cells, including the mammalian blastocyst, adrenal cortex, thyroid, anterior pituitary, and the mechanism of endoreduplication (amitotic over-replication of DNA) can be traced to lower eukaryotes. This increased activity in relation to terminal commitment is inconsistent with a simple "DNA repair' view of pol beta. It is therefore proposed that pol beta may play a more fundamental role in cellular differentiation through involvement in a putative subgenomic DNA replication-based model of terminal gene expression. Thus genetic instability, loss of differentiation, and carcinogenesis may result from aberration(s) or "derailment' of such replication-based mechanism of terminal gene expression. It is suggested to examine the relationship of DNA pol beta to genomic instability and carcinogenesis using genetic analyses and antisense technology with possible applications for gene therapy against colorectal cancer.


Assuntos
Neoplasias Colorretais/terapia , DNA Polimerase I/genética , Replicação do DNA , Terapia Genética , Neoplasias/terapia , Animais , Blastocisto/fisiologia , Ciclo Celular , Diferenciação Celular , Divisão Celular , Neoplasias Colorretais/genética , DNA Polimerase I/biossíntese , Desenvolvimento Embrionário e Fetal , Genes Supressores de Tumor , Humanos , Mamíferos , Modelos Biológicos , Mutação , Neoplasias/genética , Oncogenes
17.
Microbiol Rev ; 55(3): 512-42, 1991 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-1943999

RESUMO

The historic arguments for the participation of eukaryotic DNA replication in the control of gene expression are reconsidered along with more recent evidence. An earlier view in which gene commitment was achieved with stable chromatin structures which required DNA replication to reset expression potential (D. D. Brown, Cell 37:359-365, 1984) is further considered. The participation of nonspecific stable repressor of gene activity (histones and other chromatin proteins), as previously proposed, is reexamined. The possible function of positive trans-acting factors is now further developed by considering evidence from DNA virus models. It is proposed that these positive factors act to control the initiation of replicon-specific DNA synthesis in the S phase (early or late replication timing). Stable chromatin assembles during replication into potentially active (early S) or inactive (late S) states with prevailing trans-acting factors (early) or repressing factors (late) and may asymmetrically commit daughter templates. This suggests logical schemes for programming differentiation based on replicons and trans-acting initiators. This proposal requires that DNA replication precede major changes in gene commitment. Prior evidence against a role for DNA replication during terminal differentiation is reexamined along with other results from terminal differentiation of lower eukaryotes. This leads to a proposal that DNA replication may yet underlie terminal gene commitment, but that for it to do so there must exist two distinct modes of replication control. In one mode (mitotic replication) replicon initiation is tightly linked to the cell cycle, whereas the other mode (terminal replication) initiation is not cell cycle restricted, is replicon specific, and can lead to a terminally differentiated state. Aberrant control of mitotic and terminal modes of DNA replication may underlie the transformed state. Implications of a replicon basis for chromatin structure-function and the evolution of metazoan organisms are considered.


Assuntos
Diferenciação Celular/fisiologia , Cromatina/fisiologia , Replicação do DNA , Regulação da Expressão Gênica , Animais , Mitose , Replicon
18.
Virus Genes ; 20(3): 195-200, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-10949945

RESUMO

When introduced into EC cells of a blastocyst, polyomavirus (Py) T-Ag results in mice mosaic for T-Ag but otherwise essentially normal. It had been reported that SV40 T-Ag does not inhibit differentiation of F9 EC cells, but did inhibit endogenous retrovirus (ERV) production. We therefore sought to determine if Py T-Ag had any affect on EC derived embryoid body implantation onto mouse placenta. F9 EC cells were selected for T-Ag maintenance. Like the SV40 transformed cells, we show that these Py T-Ag selected EC cells no longer express IAP transcripts following differentiation into embryoid bodies. Normal and Py T-Ag selected F9 cells were differentiated into embryoid bodies then implanted into pseudopregnant mice. We observe, that normal F9 derived embryoid bodies underwent the initial stages of implantation whereas the Py T-Ag selected embryoid bodied did not implant. The implications of this observation with respect to trophectoderm and ERV function are discussed. We examine the idea that ERVs may be a required element for normal embryo implantation.


Assuntos
Antígenos Transformantes de Poliomavirus/metabolismo , Implantação do Embrião/fisiologia , Retrovirus Endógenos/metabolismo , Células-Tronco Neoplásicas/citologia , Animais , Blastocisto/citologia , Blastocisto/metabolismo , Diferenciação Celular , Ectoderma/citologia , Implantação do Embrião/genética , Células-Tronco de Carcinoma Embrionário , Retrovirus Endógenos/genética , Retrovirus Endógenos/crescimento & desenvolvimento , Feminino , Produtos do Gene env/metabolismo , Genes de Partícula A Intracisternal , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Células-Tronco Neoplásicas/metabolismo , Reação em Cadeia da Polimerase , RNA Mensageiro/análise , Transfecção , Células Tumorais Cultivadas
19.
Virus Genes ; 20(3): 209-16, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-10949947

RESUMO

n-Butyrate, an inhibitor of G1-to-S transition inhibits papovavirus DNA replication in cell culture. To explore the efficacy of n-butyrate in vivo and to better understand its mechanism, we studied the effect of n-butyrate on viral DNA replication in mice acutely infected with polyomavirus and in the papovavirus-infected cells in culture. Newborn mice treated with n-butyrate stop growing and become runted. When infected with polyomavirus, these mice show a strong overall inhibition of viral DNA. However, a notable exception to this was the continued viral DNA replication in the differentiated mouse keratinocytes and renal epithelial cells as determined by in situ hybridization. n-Butyrate significantly inhibited viral DNA replication in the cultured IDL cells, and in polyomavirus-infected C2C12 myoblasts based on Southern blot analysis and in situ hybridization. DNA polymerase alpha (but not DNA polymerase beta) and the characteristic nuclear expression of PCNA were both inhibited in the n-butyrate treated IDL and C2C12 cells. n-Butyrate, therefore, inhibited host and viral DNA synthesis in the undifferentiated cells.


Assuntos
Papillomavirus Bovino 1/fisiologia , Butiratos/farmacologia , Replicação do DNA , DNA Viral/biossíntese , Polyomavirus/fisiologia , Animais , Animais Recém-Nascidos , Southern Blotting , Papillomavirus Bovino 1/efeitos dos fármacos , Papillomavirus Bovino 1/metabolismo , Núcleo Celular/metabolismo , Células Cultivadas , DNA Polimerase I/antagonistas & inibidores , Hibridização In Situ , Camundongos , Camundongos Endogâmicos BALB C , Polyomavirus/efeitos dos fármacos , Polyomavirus/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo
20.
J Virol ; 74(15): 7079-84, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10888648

RESUMO

The eukaryotic replicative DNA polymerases are similar to those of large DNA viruses of eukaryotic and bacterial T4 phages but not to those of eubacteria. We develop and examine the hypothesis that DNA virus replication proteins gave rise to those of eukaryotes during evolution. We chose the DNA polymerase from phycodnavirus (which infects microalgae) as the basis of this analysis, as it represents a virus of a primitive eukaryote. We show that it has significant similarity with replicative DNA polymerases of eukaryotes and certain of their large DNA viruses. Sequence alignment confirms this similarity and establishes the presence of highly conserved domains in the polymerase amino terminus. Subsequent reconstruction of a phylogenetic tree indicates that these algal viral DNA polymerases are near the root of the clade containing all eukaryotic DNA polymerase delta members but that this clade does not contain the polymerases of other DNA viruses. We consider arguments for the polarity of this relationship and present the hypothesis that the replication genes of DNA viruses gave rise to those of eukaryotes and not the reverse direction.


Assuntos
Replicação do DNA , Vírus de DNA/genética , DNA Polimerase Dirigida por DNA/genética , Células Eucarióticas/enzimologia , Phycodnaviridae/genética , Sequência de Aminoácidos , Chlorella/virologia , Vírus de DNA/enzimologia , DNA Polimerase Dirigida por DNA/química , Evolução Molecular , Dados de Sequência Molecular , Phaeophyceae/virologia , Phycodnaviridae/enzimologia , Filogenia , Alinhamento de Sequência
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