Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:5.99.1.2 (topoisomerase)
 9,166 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The effect of the topoisomerase II inhibitor Novobiocin on T-cell mediated cytotoxicity was tested under various assay conditions. When effector cells were class I major histocompatibility complex (MHC)-specific CD8+ cytotoxic T cells (Tc), Novobiocin caused a biphasic pattern of inhibition and the two components of the inhibition could be separated based on Ca2+ requirement. Unseparated populations of class II MHC specific Tc, containing CD4+ and CD8+ effectors gave the same pattern of inhibition. When CD8+ cells were depleted from the latter population of effectors, different patterns of inhibition from those obtained with CD8+ Tc were seen and furthermore the target affected the pattern of inhibition. Overall the results add further support to there being more than one pathway of CD8+ T-cell mediated cytotoxicity and further illustrate differences between CD4+ and CD8+ T-cell mediated cytotoxicity.
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PMID:Inhibition of T-cell mediated cytotoxicity by Novobiocin suggests multiple pathways for both CD4+ and CD8+ cytotoxic T cells. 135 96

A unique transgenic mouse line has undergone transgene integration in a very precise fashion. The phenotype displayed by mice of the line followed the predicted inheritance patterns for X-linked transgene insertion which has been confirmed. In order to investigate the mechanism of integration the DNA sequence of the transgene and cellular junctions have been determined. A comparison between wild type and transgenic mutant sequences at the site of insertion revealed that there was no loss or rearrangement of cellular DNA upon integration of the transgene. The cellular sequences at the transgene 5' and 3' joins are contiguous in the wild type. The integrant exists as a head to tail tandem dimer with minimal loss of sequence compared with the injected monomer. Analysis of the site of insertion has revealed a 5 bp homology between the 5' end of the transgene and the cellular sequences. In addition, adjacent to the site of insertion within the cellular sequences, there are several sequence motifs implicated in recombination events including a clustering of strong consensus sites of DNA topoisomerase type I and a region of homology to the human minisatellite consensus core sequence, the Escherichia coli Chi site and the meiotic recombination hotspot within the E beta gene of the murine major histocompatibility complex. This clustering of features is likely to have been factorial in the integrity of the insertion event. A model depicting the mechanism of this precise integration is proposed.
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PMID:A model for the mechanism of precise integration of a microinjected transgene. 867 44

Measurement of disease activity in systemic autoimmune disorders is often unreliable, and immunosuppressive therapy is often titrated to crude clinical response and/or onset of complications. Systemic sclerosis (SSc) presents a distinct clinical phenotype associated with specific autoantibodies. Anti-topoisomerase-1 (SCL-70) is selectively detected in 30-60% of subjects with diffuse skin and interstitial lung involvement. Such patients offer an ideal clinical model to characterize and quantify the autoantigen-specific T-cell response and its correlation with disease phenotype and activity. Human leukocyte antigen A2 (HLA-A2)-restricted topo-1 peptides were selected based on an epitope prediction algorithm. For initial studies, the best binder topo-1(262-270) KMLDHEYTT (#262) was used alone or loaded onto an artificial antigen-presenting platform generated by coupling a dimeric major histocompatibility complex-immunoglobulin G fusion protein (HLA-A2-Ig) and anti-CD28 antibodies onto magnetic beads (artificial antigen-presenting cells). Blood samples (100 microL) from HLA-A2+ SSc patients and cytomegalovirus (CMV) seropositive healthy control subjects were tested in an intracellular cytokine staining assay. Gamma interferon production by CD8+ T cells was measured after stimulation with peptide #262, CMVpp65, or MART-1 (irrelevant peptide). In two of five SCL-70+ patients, peptide #262-loaded aAPCs induced a specific CD8+ T-cell response (0.45% +/- 0.23% of total CD8+ cells). This response was not observed in the seven SCL-70- (five SSc and two CMV+) control subjects studied (0.03% +/- 0.02%). Interestingly, bronchoalveolar lavage fluid obtained from one topo-1-responsive SSc patient who had worsening respiratory function and active alveolitis showed striking enrichment of topo-1-specific CD8+ T cells (3.94%). This small-volume ex vivo assay may prove to be a sensitive and specific tool to assess disease activity and to monitor response to therapy in patients with scleroderma.
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PMID:Evaluation of topoisomerase-1-specific CD8+ T-cell response in systemic sclerosis. 1646 96

Epstein-Barr virus (EBV) is tightly associated with certain human cancers, but there is as yet no specific treatment against EBV-related diseases. The EBV-encoded EBNA1 protein is essential to maintain viral episomes and for viral persistence. As such, EBNA1 is expressed in all EBV-infected cells, and is highly antigenic. All infected individuals, including individuals with cancer, have CD8(+) T cells directed towards EBNA1 epitopes, yet the immune system fails to detect and destroy cells harboring the virus. EBV immune evasion depends on the capacity of the Gly-Ala repeat (GAr) domain of EBNA1 to inhibit the translation of its own mRNA in cis, thereby limiting the production of EBNA1-derived antigenic peptides presented by the major histocompatibility complex (MHC) class I pathway. Here we establish a yeast-based assay for monitoring GAr-dependent inhibition of translation. Using this assay we identify doxorubicin (DXR) as a compound that specifically interferes with the GAr effect on translation in yeast. DXR targets the topoisomerase-II-DNA complexes and thereby causes genomic damage. We show, however, that the genotoxic effect of DXR and various analogs thereof is uncoupled from the effect on GAr-mediated translation control. This is further supported by the observation that etoposide and teniposide, representing another class of topoisomerase-II-DNA targeting drugs, have no effect on GAr-mediated translation control. DXR and active analogs stimulate, in a GAr-dependent manner, EBNA1 expression in mammalian cells and overcome GAr-dependent restriction of MHC class I antigen presentation. These results validate our approach as an effective high-throughput screening assay to identify drugs that interfere with EBV immune evasion and, thus, constitute candidates for treating EBV-related diseases, in particular EBV-associated cancers.
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PMID:A yeast-based assay identifies drugs that interfere with immune evasion of the Epstein-Barr virus. 2455 96

Recombination hotspots are the regions within the genome where the rate, and the frequency of recombination are optimum with a size varying from 1 to 2kb. The recombination event is mediated by the double-stranded break formation, guided by the combined enzymatic action of DNA topoisomerase and Spo 11 endonuclease. These regions are distributed non-uniformly throughout the human genome and cause distortions in the genetic map. Numerous lines of evidence suggest that the number of hotspots known in humans has increased manifold in recent years. A few facts about the hotspot evolutions were also put forward, indicating the differences in the hotspot position between chimpanzees and humans. In mice, recombination hot spots were found to be clustered within the major histocompatibility complex (MHC) region. Several models, that help explain meiotic recombination has been proposed. Moreover, scientists also developed some computational tools to locate the hotspot position and estimate their recombination rate in humans is of great interest to population and medical geneticists. Here we reviewed the molecular mechanisms, models and in silico prediction techniques of hot spot residues.
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PMID:Recombination hotspots: Models and tools for detection. 2699 54