ABSTRACT
Retroviral integrase catalyses the integration of viral DNA into host target DNA, which is an essential step in the life cycle of all retroviruses. Previous structural characterization of integrase-viral DNA complexes, or intasomes, from the spumavirus prototype foamy virus revealed a functional integrase tetramer, and it is generally believed that intasomes derived from other retroviral genera use tetrameric integrase. However, the intasomes of orthoretroviruses, which include all known pathogenic species, have not been characterized structurally. Here, using single-particle cryo-electron microscopy and X-ray crystallography, we determine an unexpected octameric integrase architecture for the intasome of the betaretrovirus mouse mammary tumour virus. The structure is composed of two core integrase dimers, which interact with the viral DNA ends and structurally mimic the integrase tetramer of prototype foamy virus, and two flanking integrase dimers that engage the core structure via their integrase carboxy-terminal domains. Contrary to the belief that tetrameric integrase components are sufficient to catalyse integration, the flanking integrase dimers were necessary for mouse mammary tumour virus integrase activity. The integrase octamer solves a conundrum for betaretroviruses as well as alpharetroviruses by providing critical carboxy-terminal domains to the intasome core that cannot be provided in cis because of evolutionarily restrictive catalytic core domain-carboxy-terminal domain linker regions. The octameric architecture of the intasome of mouse mammary tumour virus provides new insight into the structural basis of retroviral DNA integration.
Subject(s)
Cryoelectron Microscopy , DNA, Viral/metabolism , DNA, Viral/ultrastructure , Integrases/chemistry , Integrases/ultrastructure , Mammary Tumor Virus, Mouse/enzymology , Protein Multimerization , Catalytic Domain , Crystallography, X-Ray , DNA, Viral/chemistry , Integrases/metabolism , Mammary Tumor Virus, Mouse/chemistry , Mammary Tumor Virus, Mouse/genetics , Mammary Tumor Virus, Mouse/ultrastructure , Models, Molecular , Protein Structure, Quaternary , Spumavirus/chemistry , Spumavirus/enzymology , Virus IntegrationABSTRACT
Cryoelectron microscopy of Mouse mammary tumor virus, a Betaretrovirus, provided information about glycoprotein structure and core formation. The virions showed the broad range of diameters typical of retroviruses. Betaretroviruses assemble cytoplasmically, so the broad size range cannot reflect the use of the plasma membrane as a platform for assembly.
Subject(s)
Cryoelectron Microscopy , Mammary Tumor Virus, Mouse/ultrastructure , Cell Line , Cell Membrane/ultrastructure , Cell Membrane/virology , HIV-1/chemistry , HIV-1/growth & development , Mammary Tumor Virus, Mouse/chemistry , Mammary Tumor Virus, Mouse/growth & development , Virion/chemistry , Virion/growth & development , Virion/ultrastructure , Virus AssemblyABSTRACT
Subsaturated nucleosomal arrays were reconstituted on a single-copy MMTV promoter DNA fragment by salt dialysis procedures and studied by atomic force microscopy. Up to an occupation level of approximately eight nucleosomes on this 1900 bp template, salt reconstitution produces nucleosomal arrays which look very similar to comparably loaded 5S rDNA nucleosomal arrays; i.e., nucleosomes are dispersed on the DNA template. Thus, at these occupation levels, the single-copy MMTV template forms arrays suitable for biophysical analyses. A quantitative comparison of the population features of subsaturated MMTV and 5S arrays detects differences between the two: a requirement for higher histone levels to achieve a given level of nucleosome occupation on MMTV templates, indicating that nucleosome loading is thermodynamically less favorable on this template; a preference for pairwise nucleosome occupation of the MMTV (but not the 5S) template at midrange occupation levels; and an enhanced salt stability for nucleosomes on MMTV versus 5S arrays, particularly in the midrange of array occupation. When average occupation levels exceed approximately eight nucleosomes per template, MMTV arrays show a significant level of mainly intramolecular compaction; 5S arrays do not. Taken together, these results show clearly that the nature of the underlying DNA template can affect the physical properties of nucleosomal arrays. DNA sequence-directed differences in the physical properties of chromatin may have important consequences for functional processes such as gene regulation.
Subject(s)
Mammary Tumor Virus, Mouse/genetics , Nucleosomes/ultrastructure , Promoter Regions, Genetic , DNA/ultrastructure , Histones/metabolism , Mammary Tumor Virus, Mouse/ultrastructure , Microscopy, Atomic Force , RNA, Ribosomal, 5S/ultrastructure , Sodium Chloride/chemistry , Templates, GeneticABSTRACT
En este trabajo se describe la citología, histología y ultraestructura del tumor mamario transplantable (Ca-MMt-INHRR-984) de la cepa de ratones N: NIH (s) desarrollado a nivel intraperiritoneal y subcutáneo. Las células tumorales muestran cambios ultraestructurales relacionados con su actividad metabólica y se identificaron en el tejido signos de necrosis y degeneración en algunas células. Tanto en el tumor ascítico como en las células del tumor subcutáneo se vieron partículas virales correspodientes a retrovirus tipo B. Se discute la importancia de los hallazgos en relación con el conocimiento sobre la relación entre el MMTV y partículas similares detectadas en la leche y en las células del cáncer mamario humano, por lo que se plantea la utilidad del modelo biológico presentado para el estudio de la oncogénesis
Subject(s)
Animals , Mice , Gammaretrovirus , Mice/anatomy & histology , Models, Biological , Oncogenes , Mammary Tumor Virus, Mouse/ultrastructureABSTRACT
The current knowledge of the distribution of the mouse mammary tumor virus (MMTV) proviral genomes and the mechanism of mammary tumorigenesis by MMTV in mice, with the main emphasis on Asian feral mice, is reviewed. The relevant earlier discoveries on the mode of MMTV transmission are summarized to provide an outline of the biology of MMTV. Finally, the viral etiology of human breast cancer will be discussed.
Subject(s)
Animals, Wild/virology , Mammary Neoplasms, Experimental/virology , Mammary Tumor Virus, Mouse , Retroviridae Infections/etiology , Tumor Virus Infections/etiology , Animals , Animals, Wild/genetics , Breast Neoplasms/genetics , DNA, Neoplasm/genetics , DNA, Viral/analysis , Female , Humans , Mammary Neoplasms, Experimental/genetics , Mammary Tumor Virus, Mouse/genetics , Mammary Tumor Virus, Mouse/isolation & purification , Mammary Tumor Virus, Mouse/ultrastructure , Mice , Retroviridae Infections/genetics , Retroviridae Infections/transmission , Sequence Analysis, DNA , Tumor Virus Infections/genetics , Tumor Virus Infections/transmissionABSTRACT
Exogenous mouse mammary tumor viruses (MMTV) replicate in the mammary glands of infected females, and so infect the suckling pups. We have previously shown that the virus is rapidly disseminated to all the lymphoid organs, including the thymus. The present electron microscope immunohistochemical study describes the viral production site in the thymus. Viral buds and viral proteins were restricted to the thymus medullary epithelial cells. MMTV-encoded proteins were identified on the free viral particles and on the budding ones, the ribosomes, the membrane of the endoplasmic reticulum, and on the membrane of the medullary type II epithelial cell vacuolar network. The thymus medullary epithelial cells can thus integrate the virus and allow viral replication. The results support earlier results indicating that in some experimental conditions, epithelial cells may be involved in MMTV-induced negative selection by showing that thymic epithelial cells do express MMTV-encoded proteins.
Subject(s)
Mammary Tumor Virus, Mouse/physiology , Thymus Gland/immunology , Thymus Gland/virology , Virus Replication/immunology , Animals , Antibodies, Viral/isolation & purification , Antibody Specificity , Epithelium/immunology , Epithelium/ultrastructure , Epithelium/virology , Female , Lactation/immunology , Mammary Tumor Virus, Mouse/immunology , Mammary Tumor Virus, Mouse/ultrastructure , Mice , Mice, Inbred BALB C , Thymus Gland/ultrastructureABSTRACT
The translational positions of nucleosomes in the promoter region of the mouse mammary tumor virus (MMTV) were defined at high resolution. Nucleosome boundaries were determined in primer extension assays using full-length single-stranded mononucleosomal DNA prepared from cells treated with formaldehyde, a reversible protein-DNA cross-linking agent. Multiple boundaries were observed in both the nucleosome A (Nuc-A) and Nuc-B region of the promoter, indicating multiple nucleosome translational frames. The different nucleosome frames in both the Nuc-A and Nuc-B regions were occupied unequally. The most frequently occupied frames were found clustered within 50-60 bases of each other, resulting in a distribution centered in the positions defined previously at low resolution for Nuc-A and Nuc-B. The most abundant 5' ends of the frames in the B region were found between -235 and -187, and the 3' ends between -86 and -36, whereas in the A region the most abundant 5' ends were between -22 and +42, and the 3' ends between +121 and +186. Although frames in the Nuc-B region of the LTR extend at a low frequency in the 5' direction toward the Nuc-C region, there is a sharp discontinuity in the 3' direction toward Nuc-A, suggesting the presence of a boundary constraint in the A-B linker. The positions and relative occupancies of nucleosome frames, in either the B or the A region, did not change when the promoter was activated with dexamethasone.
Subject(s)
DNA, Viral/ultrastructure , Mammary Tumor Virus, Mouse/ultrastructure , Nucleosomes/ultrastructure , Animals , Base Sequence , Binding Sites , Cells, Cultured , Cross-Linking Reagents , DNA Primers , DNA, Single-Stranded , DNA, Viral/genetics , Macromolecular Substances , Mice , Molecular Sequence Data , Movement , Protein Binding , Repetitive Sequences, Nucleic Acid , Sequence Analysis, DNAABSTRACT
Several groups of wild mice (Mus musculus) were captured from eight different locations in Asia and bred for several generations in a facility free of any laboratory strains of mice carrying mouse mammary tumor virus (MMTV). The distribution of endogenous MMTV proviral sequences in the liver tissues of these mice was investigated by using Southern blot hybridizations. Four categories of mice were identified. Mice originating from Bogor, Indonesia (Cas-Bgr); He-mei, Taiwan (Cas-Hmi/1); and Malaysia (Cas-Mal) were found to carry an endogenous MMTV provirus consisting of the env, gag-pol, and long terminal repeat sequences. Mice captured from Kojuri, Republic of Korea (Sub-Kjr); Nagoya, Japan (Mol-nag); and three Chinese provinces, Shanghai (Sub-Shh), Beijing (Sub-Bjn), and Jiayuguang (Sub-Jyg/1), appeared to carry defective proviruses. Some mice originating from He-mei (Cas-Hmi/2) and Jiayuguang (Sub-Jyg/2) were found to be completely free of endogenous MMTV. Interestingly, however, the Sub-Jyg/2 mice, after several generations of inbreeding, were found, unlike all of the other subspecies that we examined in the present study, to develop mammary tumors at a high incidence (80 to 90%) with a short period of latency. Electron microscopic examination of the mammary glands and mammary tumors of these mice revealed the presence of numerous intracytoplasmic A, immature, budding, and mature B particles. Furthermore, the mammary tumors were found to contain MMTV proviral sequences. It seems, therefore, that Sub-Jyg/2 mice carry an exogenous MMTV which contributes to their developing mammary tumors.
Subject(s)
Animals, Wild/microbiology , Mammary Tumor Virus, Mouse/isolation & purification , Muridae/microbiology , Retroviridae Infections/veterinary , Tumor Virus Infections/veterinary , Animals , Asia, Southeastern/epidemiology , Defective Viruses/genetics , Asia, Eastern/epidemiology , Female , Genes, env , Genes, gag , Genes, pol , Genome, Viral , Mammary Neoplasms, Animal/microbiology , Mammary Neoplasms, Animal/ultrastructure , Mammary Tumor Virus, Mouse/genetics , Mammary Tumor Virus, Mouse/ultrastructure , Proviruses/genetics , Retroviridae Infections/epidemiology , Tumor Virus Infections/epidemiologyABSTRACT
The immature capsids of the mouse mammary tumor virus (MMTV), known as intracytoplasmic A particles, have been isolated from murine L1210 leukemia cells. The diameter of the isolated particles was 80 nm as determined by negative staining. Two polypeptides of 77 and 110 kDa were found to be their major polypeptide components, in agreement with the expected sizes of the Gag and Gag-Pro precursor polypeptides of the mature MMTV proteins. Both polypeptides were recognized by antibodies directed toward the matrix (p10) and capsid (p27) proteins of MMTV. Immunogold labeling of p10 on isolated A particles, visualized by negative staining, showed that this protein is located at the surface of the immature capsids, whereas p27 can be detected only in broken or disrupted particles, suggesting that it has an internal location. These observations were confirmed by immunolabeling of both proteins on thin sections of A particle-producing cells. In addition, the viral protease had a more internal position than p27. Since the sequential order of the viral proteins in the Gag precursor is p10-pp21-p27-p14 and that in Gag-Pro is p10-pp21-p27-p30-protease, our results demonstrate the radial organization of the polypeptide precursors forming the intracytoplasmic A particles.
Subject(s)
Capsid/isolation & purification , Mammary Tumor Virus, Mouse/immunology , Viral Core Proteins/isolation & purification , Amino Acid Sequence , Animals , Antigens, Viral/immunology , Capsid/immunology , Capsid/ultrastructure , Dexamethasone/pharmacology , Epitopes , Immunohistochemistry , Leukemia L1210 , Mammary Tumor Virus, Mouse/isolation & purification , Mammary Tumor Virus, Mouse/ultrastructure , Mice , Molecular Sequence Data , Tumor Cells, Cultured , Viral Core Proteins/immunology , Viral Core Proteins/ultrastructure , Virus Replication/drug effectsABSTRACT
Cleavage of oligopeptide substrates mimicking the maturation sites in the Gag polyproteins of the mouse mammary tumor virus was assayed using lysed virus. Cleavage at the expected P1-P1' positions was detected in four of seven synthetic peptides. However, studies with specific inhibitors of retroviral proteases showed that only two of them could be unequivocally attributed to the viral enzyme. In an attempt to characterize other proteolytic activities that copurify with the virus, we isolated a multicatalytic high molecular mass protease (700 kDa) that copurifies with the virus. This protein has been identified as an alpha 2-macroglobulin-protein complex according to its biochemical properties and ultrastructure. The proteases forming these complexes are mainly serine proteases and can be inhibited by phenylmethylsulfonyl fluoride. However, other compounds such as chymostatin and elastatinal are more effective inhibitors. The relative efficacy of each compound depends on the substrate, since the complexes described herein appear to be multicatalytic. Elastatinal is a very good inhibitor of the cleavages found at Ala-Ala bonds in peptides representing the capsid/nucleocapsid site, while chymostatin inhibits certain cleavages at the carboxyl terminus of bonds involving leucine and valine in three of the substrates used. Therefore, the alpha 2-macroglobulin present in the cell culture medium is able to bind proteases, forming high molecular weight complexes, which are active against peptide substrates, copurify with the virus and are responsible for the nonviral proteolytic activities found in the purified virus. Elastase appears to be the main proteolytic activity which can be detected in the alpha 2-macroglobulin-protease complexes associated with the virus.
Subject(s)
Endopeptidases/metabolism , Mammary Tumor Virus, Mouse/metabolism , alpha-Macroglobulins/metabolism , Amino Acid Sequence , Cell Line , Chromatography, Gel , Culture Media , Electrophoresis, Polyacrylamide Gel , Endopeptidases/isolation & purification , Gene Products, gag/metabolism , Hydrolysis , Mammary Tumor Virus, Mouse/enzymology , Mammary Tumor Virus, Mouse/ultrastructure , Microscopy, Electron , Molecular Sequence Data , alpha-Macroglobulins/isolation & purificationABSTRACT
Retrovirus-like particles were secreted in a steroid-dependent manner by the human mammary carcinoma cell line T47D. The particles exhibited typical retroviral properties such as their electron microscopic appearance (95 nm in diameter) and occasional budding, sedimentation at 1.14 g/ml, reverse transcriptase activity and genomic RNA. The T47D particles were related to mouse mammary tumour virus (MMTV) as shown by their ultrastructural appearance (B type-like eccentric dense cores and budding), Mg2+ dependence of the reverse transcriptase activity; immunological reactivity with MMTV-directed antibodies (revealing proteins of 63K, 52K, 26K and 18K), and hybridization of particle RNA with MMTV DNA under stringent conditions. Purified particles were able to incorporate deoxynucleoside triphosphates in the absence of an exogenous primer and template, thus indicating the existence of a complete and biochemically functional reverse transcription apparatus (reverse transcriptase, RNA and primer) and the ability to direct endogenous cDNA synthesis. Labelled particle cDNA hybridized strongly to human genomic DNA but not to mouse and cat DNA, thus indicating the human origin of the T47D particles. Furthermore all human DNAs, hybridized with the labelled particle cDNA, showed a uniform hybridization pattern of restriction fragments, indicating the endogenous origin and distribution of the proviral particle DNA in the human genome.
Subject(s)
Adenocarcinoma/microbiology , Breast Neoplasms/microbiology , Mammary Tumor Virus, Mouse/ultrastructure , Animals , Cats , Cross Reactions , DNA, Single-Stranded/biosynthesis , DNA, Viral/biosynthesis , Genome, Human , Humans , Inclusion Bodies, Viral , Mammary Tumor Virus, Mouse/immunology , Mammary Tumor Virus, Mouse/isolation & purification , Mice , Nucleic Acid Hybridization , Proviruses , RNA, Viral/analysis , RNA-Directed DNA Polymerase/analysis , Tumor Cells, CulturedABSTRACT
Fifty-two human breast tumors were screened for the presence of DNA homology to mouse mammary tumor virus (MMTV) using molecularly cloned MMTV proviral genomic DNA probes and dot-blot hybridization. Seven patients were found to contain an entire provirus (gag, pol, env, and LTR positive at high stringency). Fifty percent (5/10) of patients having a first degree relative with breast carcinoma were found to have DNA homology to the gag-pol portion of the MMTV genome when hybridization and washing was performed at moderate (56C) stringency. Thirty-nine percent (7/18) of patients with any positive family history and 23 percent (8/34) of patients with a negative family history demonstrated homology under these parameters. Of the patients positive for gag-pol at moderate stringency, fewer had taken exogenous hormones than the sample group (20 percent vs 52 percent), more were parous (93 percent vs 68 percent), estrogen receptor positive (69 percent vs 48 percent), and male (13 percent vs 4 percent). At higher stringency (62C) no correlation to family history, hormone use or sex was detected, but positivity was noted among estrogen and progesterone receptor positive patients (67 percent vs 48 percent). Under lower stringency wash conditions, mismatched MMTV-related sequences are identified suggesting the existence of an endogenous gene with partial homology to MMTV. High stringency hybridization may identify a related retrovirus with significant homology to MMTV.
Subject(s)
Breast Neoplasms/genetics , DNA, Neoplasm/analysis , DNA, Viral/analysis , Genome, Viral , Mammary Tumor Virus, Mouse/genetics , Sequence Homology, Nucleic Acid , Adult , Aged , Aged, 80 and over , Animals , Breast Neoplasms/chemistry , Breast Neoplasms/pathology , Estrogens/pharmacology , Female , Genes, gag , Genes, pol , Humans , Male , Mammary Tumor Virus, Mouse/ultrastructure , Mice , Mice, Inbred C3H , Middle Aged , Progesterone/pharmacology , Proviruses/genetics , Reference Standards , Sex FactorsABSTRACT
DNA sequences from the long terminal repeat of the mouse mammary tumor virus (MMTV-LTR) position nucleosomes both in vivo and in vitro. Here, were present chromatin reconstitution experiments showing that MMTV-LTR sequences from -236 to +204 accommodate two histone octamers in positions compatible with the in vivo data. This positioning is not influenced by the length of the DNA fragment and occurs in linear as well as in closed circular DNA molecules. MMTV-LTR DNA sequences show an intrinsic bendability that closely resembles its wrapping around the histone octamer. We propose that bendability is responsible for the observed rotational nucleosome positioning. Translational nucleosome positioning seems also to be determined by the DNA sequence. These data, along with the results from reconstitution experiments with insertion mutants, support a modular model of nucleosome phasing on MMTV-LTR, where the actual positioning of the histone octamer results from the additive effect of multiple features of the DNA sequence.
Subject(s)
Mammary Tumor Virus, Mouse/ultrastructure , Nucleosomes/ultrastructure , Regulatory Sequences, Nucleic Acid , Repetitive Sequences, Nucleic Acid , Base Sequence , DNA, Circular/ultrastructure , DNA, Viral/ultrastructure , Deoxyribonuclease I/pharmacology , In Vitro Techniques , Molecular Sequence DataABSTRACT
Exogenous mouse mammary tumor virus (MMTV) was cloned from a GR mammary tumor. Clone lambda GRT39 contained a full-length integrated MMTV(GR) provirus and both 5' and 3' host flanking DNA. The lambda GRT39 provirus had no apparent structural changes associated with cloning and retained the exogenous MMTV gag gene poison sequence. When introduced into rat mammary adenocarcinoma LA7 cells, the lambda GRT39 provirus was fully expressed. lambda GRT39-transfected LA7 cells made MMTV RNA, had gp52 SU protein on the cell surface, and produced B-type retrovirus particles characteristic of MMTV. Mammary tumors developed in hormone-stimulated BALB/c females injected with MMTV from lambda GRT39-transfected LA7 cells [MMTV (lambda GRT39)]. The tumors had new, clonally integrated copies of the MMTV(lambda GRT39) provirus and were expressing MMTV antigen. These data indicate that the lambda GRT39 provirus is biologically active and pathogenic.
Subject(s)
Adenocarcinoma/microbiology , Mammary Neoplasms, Experimental/microbiology , Mammary Tumor Virus, Mouse/pathogenicity , Animals , Animals, Newborn , Blotting, Southern , Cloning, Molecular , DNA, Viral/analysis , DNA, Viral/genetics , Female , Gene Expression Regulation , Mammary Tumor Virus, Mouse/genetics , Mammary Tumor Virus, Mouse/ultrastructure , Mice , Mice, Inbred BALB C , Microscopy, Electron , Proviruses/genetics , Proviruses/pathogenicity , Rats , Restriction Mapping , Tumor Cells, CulturedABSTRACT
The retrovirus mouse mammary tumor virus is present in mouse strains with a high incidence of mammary tumors as a causative agent. It is produced mainly in the mammary glands of sexually mature females and is milk-transmitted to newborns. The fate of the mouse mammary tumor virus is almost unknown. Where it enters, how it is distributed, and where it remains latent, remain unresolved problems. This study tries to answer the first of these questions. Viruses are for the most part digested in the stomach. Very few well-preserved B particles, i.e., the infective particles, are allowed to enter through a process of endocytosis, mainly in the newborn-type epithelial cells. These are epithelial cells with a very rich absorptive apparatus, characteristic of newborn rodents. The adult-type absorptive cells and the M cells of the Peyer's patches might be partly involved.
Subject(s)
Animals, Newborn/microbiology , Intestinal Absorption , Intestine, Small/microbiology , Mammary Tumor Virus, Mouse/ultrastructure , Animals , Intestine, Small/ultrastructure , Mice , Mice, Inbred BALB C , Virion/ultrastructureABSTRACT
S49 mouse lymphoma cells were found to be extremely sensitive to the antiproliferative activity of interferon. These characteristics were studied to select for IFN-resistant cell variants. Some 0.6% of the parental S49 cell population were resistant to the antiproliferative and cytotoxic activities of IFN. The resistant cells were cloned and analyzed for their responses to several of the activities of IFN, namely, inhibition of encephalomyocarditis (EMC) virus, murine leukemia virus (MuLV) replications, and the induction of (2'-5') oligoadenylate synthetase. Among the clones selected some were highly resistant while others demonstrated only partial responsiveness to IFN. S49 cells demonstrate tubular structures in the cytoplasm. These structures were previously reported to be antigenically related to mouse mammary tumor virus (MMTV). We report here that IFN treatment decreases the expression of these cytoplasmic viral structures as revealed by electron microscopy. To correlate this novel antiviral activity to the more established functions of IFN we utilized the above mentioned S49 IFN-resistant variants. The anti-MMTV activity of IFN correlated with the other effects of IFN in both the highly resistant and partially responsive S49 clones. Our findings indicate that a relatively high proportion of S49 cells vary in their response to IFN. The defect in the resistant cells appears to affect a primary response to IFN which is common to its diverse activities. Furthermore, the effect of IFN on MMTV-related structures involves the usual pathway of IFN action.
Subject(s)
Interferons/pharmacology , Lymphoma/pathology , 2',5'-Oligoadenylate Synthetase/biosynthesis , Animals , Cell Division , Clone Cells , Cytoplasm/microbiology , Drug Resistance , Encephalomyocarditis virus/physiology , Enzyme Induction , Leukemia Virus, Murine/physiology , Lymphoma/microbiology , Mammary Tumor Virus, Mouse/ultrastructure , Mice , Microscopy, Electron , Tumor Cells, Cultured , Virus ReplicationSubject(s)
Antigens, Viral/analysis , Immunohistochemistry , Mammary Tumor Virus, Mouse/ultrastructure , Animals , Antibodies, Monoclonal , Antibodies, Viral , Female , Mammary Neoplasms, Experimental/microbiology , Mammary Neoplasms, Experimental/ultrastructure , Mammary Tumor Virus, Mouse/immunology , Mice , Mice, Inbred C3H , Microscopy, Electron , Viral Proteins/analysis , Viral Proteins/immunologyABSTRACT
Diferentes antigenos del virus productor de tumores mamarios del raton (MMTV) fueron detectados con la aplicación de varios anticuerpos poli monoespecíficos y una técnica inmunocitoquímica de alta resolución con oro coloidal. Anticuerpos preparados contra el virus total aislado (partículas B), las proteínas p14, p25 y una glicoproteína gp55 fueron marcados con el complejo oro coloidal proteína A en secciones de carcinomas mamarios espontáneos del raton, incluidos en resina acrílica (L. R. Whrite, London Resin Company). La estructura proteica de las nucleocapsides y la envoltura viral fueron los componentes más inmunoreactivos de la subestrutura del virus MMTV. Una continuidad de los constituyentes antigénicos del virus fueron encontrados en las diferentes etapas de la morfogénesis del virus demonstrándose una correlación entre estructuras precursoras y el virus infeccioso (AU)
Subject(s)
Mice , Animals , Female , Mammary Tumor Virus, Mouse/ultrastructure , Mammary Neoplasms, Experimental/ultrastructure , Antigens, Viral, Tumor/isolation & purification , Mammary Neoplasms, Experimental/immunologyABSTRACT
Diferentes antigenos del virus productor de tumores mamarios del raton (MMTV) fueron detectados con la aplicación de varios anticuerpos poli monoespecíficos y una técnica inmunocitoquímica de alta resolución con oro coloidal. Anticuerpos preparados contra el virus total aislado (partículas B), las proteínas p14, p25 y una glicoproteína gp55 fueron marcados con el complejo oro coloidal proteína A en secciones de carcinomas mamarios espontáneos del raton, incluidos en resina acrílica (L. R. Whrite, London Resin Company). La estructura proteica de las nucleocapsides y la envoltura viral fueron los componentes más inmunoreactivos de la subestrutura del virus MMTV. Una continuidad de los constituyentes antigénicos del virus fueron encontrados en las diferentes etapas de la morfogénesis del virus demonstrándose una correlación entre estructuras precursoras y el virus infeccioso