Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 74
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Int J Mol Sci ; 22(2)2021 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-33477564

RESUMO

Deoxyribonucleic acid (DNA) replication can be divided into three major steps: initiation, elongation and termination. Each time a human cell divides, these steps must be reiteratively carried out. Disruption of DNA replication can lead to genomic instability, with the accumulation of point mutations or larger chromosomal anomalies such as rearrangements. While cancer is the most common class of disease associated with genomic instability, several congenital diseases with dysfunctional DNA replication give rise to similar DNA alterations. In this review, we discuss all congenital diseases that arise from pathogenic variants in essential replication genes across the spectrum of aberrant replisome assembly, origin activation and DNA synthesis. For each of these conditions, we describe their clinical phenotypes as well as molecular studies aimed at determining the functional mechanisms of disease, including the assessment of genomic stability. By comparing and contrasting these diseases, we hope to illuminate how the disruption of DNA replication at distinct steps affects human health in a surprisingly cell-type-specific manner.


Assuntos
Replicação do DNA/genética , Instabilidade Genômica/genética , Neoplasias/genética , RecQ Helicases/genética , Craniossinostoses/genética , Craniossinostoses/imunologia , Replicação do DNA/imunologia , Instabilidade Genômica/imunologia , Humanos , Mutação/genética , Neoplasias/epidemiologia , Fenótipo , RecQ Helicases/imunologia
2.
Biol Pharm Bull ; 43(11): 1715-1728, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33132317

RESUMO

SV40-encoded microRNA (miRNA), miR-S1, downregulates the large and small T antigens (LTag and STag), which promote viral replication and cellular transformation, thereby presumably impairing LTag and STag functions essential for the viral life cycle. To explore the functional significance of miR-S1-mediated downregulation of LTag and STag as well as the functional roles of miR-S1, we evaluated viral DNA replication and proinflammatory cytokine induction in cells transfected with simian virus 40 (SV40) genome plasmid and its mutated form lacking miR-S1 expression. The SV40 genome encodes two mature miR-S1s, miR-S1-3p and miR-S1-5p, of which miR-S1-3p is the predominantly expressed form. MiR-S1-3p exerted strong repressive effects on a reporter containing full-length sequence complementarity, but only marginal effect on one harboring a sequence complementary to its seed sequence. Consistently, miR-S1-3p downregulated LTag and STag transcripts with complete sequence complementarity through miR-S1-3p-Ago2-mediated mRNA decay. Transfection of SV40 plasmid induced higher DNA replication and lower LTag and STag transcripts in most of the examined cells compared to that miR-S1-deficient SV40 plasmid. However, miR-S1 itself did not affect DNA replication without the downregulation of LTag transcripts. Both LTag and STag induced the expression of tumor necrosis factor α (TNFα) and interleukin (IL)-17F, which was slightly reduced by miR-S1 due to miR-S1-mediated downregulation of LTag and STag. Forced miR-S1 expression did not affect TNFα expression, but increased IL-17F expression. Overall, our findings suggest that miR-S1-3p is a latent modifier of LTag and STag functions, ensuring efficient viral replication and attenuating cytokine expression detrimental to the viral life cycle.


Assuntos
Antígenos Virais de Tumores/genética , Regulação Viral da Expressão Gênica/imunologia , MicroRNAs/metabolismo , RNA Viral/metabolismo , Vírus 40 dos Símios/genética , Células A549 , Replicação do DNA/imunologia , DNA Viral/biossíntese , Células HEK293 , Interações entre Hospedeiro e Microrganismos/genética , Interações entre Hospedeiro e Microrganismos/imunologia , Humanos , Interleucina-17/metabolismo , Interleucina-8/metabolismo , Infecções por Polyomavirus/genética , Infecções por Polyomavirus/imunologia , Infecções por Polyomavirus/virologia , Vírus 40 dos Símios/imunologia , Fator de Necrose Tumoral alfa/metabolismo , Infecções Tumorais por Vírus/genética , Infecções Tumorais por Vírus/imunologia , Infecções Tumorais por Vírus/virologia , Replicação Viral/imunologia
3.
Cell Death Differ ; 27(11): 3162-3176, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32494026

RESUMO

ATR is a master regulator of cell response to replication stress. Adequate activation of ATR is essential for preventing genome aberrance induced by replication defect. However, the mechanism underlying ATR activation is not fully understood. Here, we identify that RBMX is an ssDNA binding protein that orchestrates a novel pathway to activate ATR. Using super-resolution STORM, we observe that RBMX and RPA bind to adjacent but nonoverlapping sites on ssDNA in response to replication stress. RBMX then binds to and facilitates positioning of TopBP1, which activates nearby ATR associated with RPA. In addition, ATR activation by ssDNA-RBMX-TopBP1 is independent of ssDNA-dsDNA junction and 9-1-1 complex. ChIP-seq analysis reveals that RBMX/RPA are highly enriched on repetitive DNAs, which are considered as fragile sites with high replication stress. RBMX depletion leads to defective localization of TopBP1 to replication stressed sites and inadequate activation of ATR. Furthermore, cells with deficient RBMX demonstrate replication defect, leading to formation of micronuclei and a high rate of sister-chromatin exchange, indicative of genome instability. Together, the results identify a new ssDNA-RBMX-TopBP1 pathway that is specifically required for activation of ATR on repetitive DNAs. Therefore, RBMX is a key factor to ensure genome stability during replication.


Assuntos
Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , Proteínas de Transporte/metabolismo , DNA de Cadeia Simples/metabolismo , Proteínas de Ligação a DNA/metabolismo , Ribonucleoproteínas Nucleares Heterogêneas/metabolismo , Proteínas Nucleares/metabolismo , Proteínas Mutadas de Ataxia Telangiectasia/genética , Cromatina/metabolismo , Replicação do DNA/imunologia , Instabilidade Genômica , Células HEK293 , Células HeLa , Ribonucleoproteínas Nucleares Heterogêneas/genética , Humanos , Fosforilação , Transdução de Sinais
4.
J Immunol ; 203(6): 1457-1467, 2019 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-31391233

RESUMO

IL-33 is an IL-1 family member protein that is a potent driver of inflammatory responses in both allergic and nonallergic disease. This proinflammatory effect is mediated primarily by extracellular release of IL-33 from stromal cells and binding of the C-terminal domain of IL-33 to its receptor ST2 on targets such as CD4+ Th2 cells, ILC2, and mast cells. Notably, IL-33 has a distinct N-terminal domain that mediates nuclear localization and chromatin binding. However, a defined in vivo cell-intrinsic role for IL-33 has not been established. We identified IL-33 expression in the nucleus of progenitor B (pro-B) and large precursor B cells in the bone marrow, an expression pattern unique to B cells among developing lymphocytes. The IL-33 receptor ST2 was not expressed within the developing B cell lineage at either the transcript or protein level. RNA sequencing analysis of wild-type and IL-33-deficient pro-B and large precursor B cells revealed a unique, IL-33-dependent transcriptional profile wherein IL-33 deficiency led to an increase in E2F targets, cell cycle genes, and DNA replication and a decrease in the p53 pathway. Using mixed bone marrow chimeric mice, we demonstrated that IL-33 deficiency resulted in an increased frequency of developing B cells via a cell-intrinsic mechanism starting at the pro-B cell stage paralleling IL-33 expression. Finally, IL-33 was detectable during early B cell development in humans and IL33 mRNA expression was decreased in B cell chronic lymphocytic leukemia samples compared with healthy controls. Collectively, these data establish a cell-intrinsic, ST2-independent role for IL-33 in early B cell development.


Assuntos
Linfócitos B/imunologia , Interleucina-33/imunologia , Adulto , Animais , Replicação do DNA/imunologia , Feminino , Humanos , Leucemia Linfocítica Crônica de Células B/imunologia , Masculino , Mastócitos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , RNA Mensageiro/imunologia , Transdução de Sinais/imunologia , Células Th2/imunologia , Proteína Supressora de Tumor p53/imunologia
5.
Nat Commun ; 10(1): 3023, 2019 07 09.
Artigo em Inglês | MEDLINE | ID: mdl-31289266

RESUMO

The largest gene families in eukaryotes are subject to allelic exclusion, but mechanisms underpinning single allele selection and inheritance remain unclear. Here, we describe a protein complex sustaining variant surface glycoprotein (VSG) allelic exclusion and antigenic variation in Trypanosoma brucei parasites. The VSG-exclusion-1 (VEX1) protein binds both telomeric VSG-associated chromatin and VEX2, an ortholog of nonsense-mediated-decay helicase, UPF1. VEX1 and VEX2 assemble in an RNA polymerase-I transcription-dependent manner and sustain the active, subtelomeric VSG-associated transcription compartment. VSG transcripts and VSG coats become highly heterogeneous when VEX proteins are depleted. Further, the DNA replication-associated chromatin assembly factor, CAF-1, binds to and specifically maintains VEX1 compartmentalisation following DNA replication. Thus, the VEX-complex controls VSG-exclusion, while CAF-1 sustains VEX-complex inheritance in association with the active-VSG. Notably, the VEX2-orthologue and CAF-1 in mammals are also implicated in exclusion and inheritance functions. In trypanosomes, these factors sustain a highly effective and paradigmatic immune evasion strategy.


Assuntos
Variação Antigênica/genética , Epigênese Genética/imunologia , Proteínas de Protozoários/genética , Trypanosoma brucei brucei/genética , Glicoproteínas Variantes de Superfície de Trypanosoma/genética , Alelos , Animais , Variação Antigênica/imunologia , Linhagem Celular , Fator 1 de Modelagem da Cromatina/imunologia , Fator 1 de Modelagem da Cromatina/metabolismo , Replicação do DNA/imunologia , Regulação da Expressão Gênica/imunologia , Interações Hospedeiro-Parasita/genética , Interações Hospedeiro-Parasita/imunologia , Evasão da Resposta Imune , Proteínas de Protozoários/imunologia , Transcrição Gênica/imunologia , Trypanosoma brucei brucei/imunologia , Tripanossomíase Africana/imunologia , Tripanossomíase Africana/parasitologia , Glicoproteínas Variantes de Superfície de Trypanosoma/imunologia , Glicoproteínas Variantes de Superfície de Trypanosoma/metabolismo
6.
Cell Microbiol ; 21(10): e13080, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31265755

RESUMO

Brucella melitensis infection causes acute necrotizing inflammation in pregnant animals; however, the pathophysiological mechanisms leading to placentitis are unknown. Here, we demonstrate that high-mobility group box 1 (HMGB1) acts as a mediator of placenta inflammation in B. melitensis-infected pregnant mice model. HMGB1 levels were increased in trophoblasts or placental explant during B. melitensis infection. Inhibition of HMGB1 activity with neutralising antibody significantly reduced the secretion of inflammatory cytokines in B. melitensis-infected trophoblasts or placenta, whereas administration of recombinant HMGB1 (rHMGB1) increased the inflammatory response. Mechanistically, this decreased inflammatory response results from inhibition of HMGB1 activity, which cause the suppression of both mitogen-activated protein kinases and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation. Moreover, neutralising antibody to HMGB1 prevented B. melitensis infection-induced activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in trophoblasts. In contrast, in vitro stimulation of trophoblasts with rHMGB1 caused activation of NADPH oxidase and increased the production of ROS, which contributes to high bacterial burden within trophoblasts or placenta. In vivo, treatment with anti-HMGB1 antibody increases the number of Brucella survival within placenta in B. melitensis-infected pregnant mice but successfully reduced the severity of placentitis and abortion.


Assuntos
Brucella melitensis/fisiologia , Brucelose/imunologia , Proteína HMGB1/metabolismo , Placenta/imunologia , Trofoblastos/metabolismo , Trofoblastos/microbiologia , Aborto Espontâneo/genética , Aborto Espontâneo/metabolismo , Aborto Espontâneo/microbiologia , Animais , Brucella melitensis/genética , Brucella melitensis/metabolismo , Brucella melitensis/patogenicidade , Brucelose/genética , Brucelose/metabolismo , Citocinas/metabolismo , Replicação do DNA/imunologia , Feminino , Proteína HMGB1/administração & dosagem , Proteína HMGB1/antagonistas & inibidores , Proteína HMGB1/genética , Inflamação/imunologia , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NADPH Oxidases/química , NADPH Oxidases/genética , NADPH Oxidases/metabolismo , Fosforilação , Placenta/microbiologia , Placenta/patologia , Gravidez , Espécies Reativas de Oxigênio/metabolismo , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Trofoblastos/enzimologia
7.
Nat Commun ; 10(1): 1688, 2019 04 11.
Artigo em Inglês | MEDLINE | ID: mdl-30975999

RESUMO

Archaea and Bacteria have evolved different defence strategies that target virtually all steps of the viral life cycle. The diversified virion morphotypes and genome contents of archaeal viruses result in a highly complex array of archaea-virus interactions. However, our understanding of archaeal antiviral activities lags far behind our knowledges of those in bacteria. Here we report a new archaeal defence system that involves DndCDEA-specific DNA phosphorothioate (PT) modification and the PbeABCD-mediated halt of virus propagation via inhibition of DNA replication. In contrast to the breakage of invasive DNA by DndFGH in bacteria, DndCDEA-PbeABCD does not degrade or cleave viral DNA. The PbeABCD-mediated PT defence system is widespread and exhibits extensive interdomain and intradomain gene transfer events. Our results suggest that DndCDEA-PbeABCD is a new type of PT-based virus resistance system, expanding the known arsenal of defence systems as well as our understanding of host-virus interactions.


Assuntos
Archaea/fisiologia , Proteínas Arqueais/metabolismo , Vírus de Archaea/genética , DNA Viral/metabolismo , Interações entre Hospedeiro e Microrganismos/genética , Archaea/virologia , Proteínas Arqueais/genética , Proteínas Arqueais/imunologia , Vírus de Archaea/patogenicidade , Replicação do DNA/imunologia , Transferência Genética Horizontal/imunologia , Imunidade Inata/genética , Imunidade Inata/imunologia , Oligonucleotídeos Fosforotioatos/metabolismo , RNA Arqueal/genética , RNA Arqueal/isolamento & purificação , Análise de Sequência de DNA
8.
Cell Cycle ; 18(8): 898-913, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30975033

RESUMO

Besides TopBP1, ETAA1 has been identified more recently as an activator of the ATR-ATRIP complex in human cells. We have examined the role of ETAA1 in the Xenopus egg-extract system, which has been instrumental in the study of ATR-ATRIP. Depletion of ETAA1 from egg extracts did not noticeably reduce the activation of ATR-ATRIP in response to replication stress, as monitored by the ATR-dependent phosphorylation of Chk1 and RPA. Moreover, lack of ETAA1 did not appear to affect DNA replication during an unperturbed S-phase. Significantly, we find that TopBP1 is considerably more abundant than ETAA1 in egg extracts. We proceeded to show that ETAA1 could support the activation of ATR-ATRIP in response to replication stress if we increased its concentration in egg extracts by adding extra full-length recombinant ETAA1. Thus, TopBP1 appears to be the predominant activator of ATR-ATRIP in response to replication stress in this system. We have also explored the biochemical mechanism by which ETAA1 activates ATR-ATRIP. We have developed an in vitro system in which full-length recombinant ETAA1 supports activation of ATR-ATRIP in the presence of defined components. We find that binding of ETAA1 to RPA associated with single-stranded DNA (ssDNA) greatly stimulates its ability to activate ATR-ATRIP. Thus, RPA-coated ssDNA serves as a direct positive effector in the ETAA1-mediated activation of ATR-ATRIP.


Assuntos
Antígenos de Superfície/metabolismo , DNA de Cadeia Simples/metabolismo , Proteína de Replicação A/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Animais , Anticorpos/imunologia , Antígenos de Superfície/imunologia , Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , Quinase 1 do Ponto de Checagem/metabolismo , Cromatina/metabolismo , Replicação do DNA/imunologia , Proteínas de Ligação a DNA/metabolismo , Fosforilação/imunologia , Ligação Proteica , Proteínas Recombinantes/metabolismo , Fase S/imunologia , Xenopus , Proteínas de Xenopus/metabolismo
9.
PLoS Pathog ; 14(11): e1007321, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30440029

RESUMO

Antigenic variation by variant surface glycoprotein (VSG) coat switching in African trypanosomes is one of the most elaborate immune evasion strategies found among pathogens. Changes in the identity of the transcribed VSG gene, which is always flanked by 70-bp and telomeric repeats, can be achieved either by transcriptional or DNA recombination mechanisms. The major route of VSG switching is DNA recombination, which occurs in the bloodstream VSG expression site (ES), a multigenic site transcribed by RNA polymerase I. Recombinogenic VSG switching is frequently catalyzed by homologous recombination (HR), a reaction normally triggered by DNA breaks. However, a clear understanding of how such breaks arise-including whether there is a dedicated and ES-focused mechanism-is lacking. Here, we synthesize data emerging from recent studies that have proposed a range of mechanisms that could generate these breaks: action of a nuclease or nucleases; repetitive DNA, most notably the 70-bp repeats, providing an intra-ES source of instability; DNA breaks derived from the VSG-adjacent telomere; DNA breaks arising from high transcription levels at the active ES; and DNA lesions arising from replication-transcription conflicts in the ES. We discuss the evidence that underpins these switch-initiation models and consider what features and mechanisms might be shared or might allow the models to be tested further. Evaluation of all these models highlights that we still have much to learn about the earliest acting step in VSG switching, which may have the greatest potential for therapeutic intervention in order to undermine the key reaction used by trypanosomes for their survival and propagation in the mammalian host.


Assuntos
Trypanosoma/imunologia , Glicoproteínas Variantes de Superfície de Trypanosoma/genética , Glicoproteínas Variantes de Superfície de Trypanosoma/imunologia , Variação Antigênica/genética , Variação Antigênica/fisiologia , DNA/metabolismo , Replicação do DNA/imunologia , Evasão da Resposta Imune/genética , Evasão da Resposta Imune/imunologia , Telômero/genética , Transcrição Gênica/genética , Trypanosoma/genética , Trypanosoma brucei brucei/metabolismo , Tripanossomíase Africana/genética , Tripanossomíase Africana/imunologia
10.
J Virol ; 92(7)2018 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-29343579

RESUMO

Genome uncoating is essential for replication of most viruses. For poxviruses, the process is divided into two stages: removal of the envelope, allowing early gene expression, and breaching of the core wall, allowing DNA release, replication, and late gene expression. Subsequent studies showed that the host proteasome and the viral D5 protein, which has an essential role in DNA replication, are required for vaccinia virus (VACV) genome uncoating. In a search for additional VACV uncoating proteins, we noted a report that described a defect in DNA replication and late expression when the gene encoding a 68-kDa ankyrin repeat/F-box protein (68k-ank), associated with the cellular SCF (Skp1, cullin1, F-box-containing complex) ubiquitin ligase complex, was deleted from the attenuated modified vaccinia virus Ankara (MVA). Here we showed that the 68k-ank deletion mutant exhibited diminished genome uncoating, formation of DNA prereplication sites, and degradation of viral cores as well as an additional, independent defect in DNA synthesis. Deletion of the 68k-ank homolog of VACV strain WR, however, was without effect, suggesting the existence of compensating genes. By inserting VACV genes into an MVA 68k-ank deletion mutant, we discovered that M2, a member of the poxvirus immune evasion (PIE) domain superfamily and a regulator of NF-κB, and C5, a member of the BTB/Kelch superfamily associated with cullin-3-based ligase complexes, independently rescued the 68k-ank deletion phenotype. Thus, poxvirus uncoating and DNA replication are intertwined processes involving at least three viral proteins with mutually redundant functions in addition to D5.IMPORTANCE Poxviruses comprise a family of large DNA viruses that infect vertebrates and invertebrates and cause diseases of medical and zoological importance. Poxviruses, unlike most other DNA viruses, replicate in the cytoplasm, and their large genomes usually encode 200 or more proteins with diverse functions. About 90 genes may be essential for chordopoxvirus replication based either on their conservation or individual gene deletion studies. However, this number may underestimate the true number of essential functions because of redundancy. Here we show that any one of three seemingly unrelated and individually nonessential proteins is required for the incompletely understood processes of genome uncoating and DNA replication, an example of synthetic lethality. Thus, poxviruses appear to have a complex genetic interaction network that has not been fully appreciated and which will require multifactor deletion screens to assess.


Assuntos
Proteínas Culina/imunologia , Replicação do DNA/imunologia , DNA Viral/imunologia , Genoma Viral/imunologia , Evasão da Resposta Imune , Proteínas Quinases Associadas a Fase S/imunologia , Vaccinia virus/imunologia , Proteínas Culina/genética , DNA Viral/genética , Células HeLa , Humanos , Proteínas Quinases Associadas a Fase S/genética , Vaccinia virus/genética
11.
Mol Immunol ; 91: 8-16, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28858629

RESUMO

B cell activation is dependent on a large increase in transcriptional output followed by focused expression on secreted immunoglobulin as the cell transitions to an antibody producing plasma cell. The rapid transcriptional induction is facilitated by the release of poised RNA pol II into productive elongation through assembly of the super elongation complex (SEC). We report that a SEC component, the Eleven -nineteen Lysine-rich leukemia (ELL) family member 3 (ELL3) is dynamically up-regulated in mature and activated human B cells followed by suppression as B cells transition to plasma cells in part mediated by the transcription repressor PRDM1. Burkitt's lymphoma and a sub-set of Diffuse Large B cell lymphoma cell lines abundantly express ELL3. Depletion of ELL3 in the germinal center derived lymphomas results in severe disruption of DNA replication and cell division along with increased DNA damage and cell death. This restricted utilization and survival dependence reveal a key step in B cell activation and indicate a potential therapeutic target against B cell lymphoma's with a germinal center origin.


Assuntos
Linfócitos B/imunologia , Divisão Celular/imunologia , Regulação da Expressão Gênica/imunologia , Fatores de Elongação da Transcrição/imunologia , Divisão Celular/genética , Sobrevivência Celular/genética , Sobrevivência Celular/imunologia , Replicação do DNA/genética , Replicação do DNA/imunologia , Humanos , Células Jurkat , RNA Polimerase II/genética , RNA Polimerase II/imunologia , Fatores de Elongação da Transcrição/genética
12.
J Clin Invest ; 127(5): 1616-1617, 2017 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-28414291

RESUMO

Patients who present with unique immunological phenotypes provide an opportunity to better understand defect-driving mutations. In this issue of the JCI, Cottineau and colleagues characterize 5 individuals who exhibited growth restriction, facial deformities, and a history of bacterial and viral infection. Further characterization revealed that these patients were neutropenic and NK cell deficient. These phenotypes were unexpectedly linked to mutations in the gene encoding a subunit of the Go-Ichi-Ni-San (GINS) complex, which is essential for DNA replication prior to cell division. Together, the results of this study lay the groundwork for future studies to explore the role of DNA replication in immune cell generation and function.


Assuntos
Anormalidades Múltiplas , Infecções Bacterianas , Replicação do DNA , Células Matadoras Naturais/imunologia , Mutação , Neutropenia , Neutrófilos/imunologia , Viroses , Anormalidades Múltiplas/genética , Anormalidades Múltiplas/imunologia , Anormalidades Múltiplas/patologia , Animais , Infecções Bacterianas/genética , Infecções Bacterianas/imunologia , Infecções Bacterianas/patologia , Divisão Celular/genética , Divisão Celular/imunologia , Replicação do DNA/genética , Replicação do DNA/imunologia , Proteínas de Ligação a DNA/imunologia , Humanos , Células Matadoras Naturais/patologia , Neutropenia/genética , Neutropenia/imunologia , Neutropenia/patologia , Neutrófilos/patologia , Viroses/genética , Viroses/imunologia , Viroses/patologia
13.
Cell Rep ; 18(5): 1312-1323, 2017 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-28147283

RESUMO

How type I and II interferons prevent periodic reemergence of latent pathogens in tissues of diverse cell types remains unknown. Using homogeneous neuron cultures latently infected with herpes simplex virus 1, we show that extrinsic type I or II interferon acts directly on neurons to induce unique gene expression signatures and inhibit the reactivation-specific burst of viral genome-wide transcription called phase I. Surprisingly, interferons suppressed reactivation only during a limited period early in phase I preceding productive virus growth. Sensitivity to type II interferon was selectively lost if viral ICP0, which normally accumulates later in phase I, was expressed before reactivation. Thus, interferons suppress reactivation by preventing initial expression of latent genomes but are ineffective once phase I viral proteins accumulate, limiting interferon action. This demonstrates that inducible reactivation from latency is only transiently sensitive to interferon. Moreover, it illustrates how latent pathogens escape host immune control to periodically replicate by rapidly deploying an interferon-resistant state.


Assuntos
Replicação do DNA/imunologia , Expressão Gênica/imunologia , Latência Viral/imunologia , Replicação Viral/imunologia , Animais , Células Cultivadas , Genoma Viral/imunologia , Herpesvirus Humano 1/imunologia , Interferon Tipo I/imunologia , Interferon gama/imunologia , Neurônios/imunologia , Neurônios/virologia , Ratos , Ratos Sprague-Dawley , Transcrição Gênica/imunologia , Proteínas Virais/imunologia , Ativação Viral/imunologia
14.
Cell Rep ; 17(11): 2927-2942, 2016 12 13.
Artigo em Inglês | MEDLINE | ID: mdl-27974207

RESUMO

Class switch recombination (CSR) at the immunoglobulin heavy chain (IgH) locus generates antibody isotypes. CSR depends on double-strand breaks (DSBs) induced by activation-induced cytidine deaminase (AID). Although DSB formation and repair machineries are active in G1 phase, efficient CSR is dependent on cell proliferation and S phase entry; however, the underlying mechanisms are obscure. Here, we show that efficient CSR requires the replicative helicase, the Mcm complex. Mcm proteins are enriched at IgH switch regions during CSR, leading to assembly of facultative replication origins that require Mcm helicase function for productive CSR. Assembly of CSR-associated origins is facilitated by R loops and promotes the physical proximity (synapsis) of recombining switch regions, which is reduced by R loop inhibition or Mcm complex depletion. Thus, R loops contribute to replication origin specification that promotes DSB resolution in CSR. This suggests a mechanism for the dependence of CSR on S phase and cell division.


Assuntos
Citidina Desaminase/genética , Replicação do DNA/imunologia , Switching de Imunoglobulina/imunologia , Cadeias Pesadas de Imunoglobulinas/imunologia , Proteínas de Manutenção de Minicromossomo/genética , Animais , Anticorpos/genética , Anticorpos/imunologia , Citidina Desaminase/imunologia , Quebras de DNA de Cadeia Dupla , Reparo do DNA/genética , Reparo do DNA/imunologia , Replicação do DNA/genética , Switching de Imunoglobulina/genética , Cadeias Pesadas de Imunoglobulinas/genética , Camundongos , Proteínas de Manutenção de Minicromossomo/imunologia , Origem de Replicação/genética
15.
Science ; 349(6248): 643-6, 2015 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-26184917

RESUMO

The germinal center (GC) is a microanatomical compartment wherein high-affinity antibody-producing B cells are selectively expanded. B cells proliferate and mutate their antibody genes in the dark zone (DZ) of the GC and are then selected by T cells in the light zone (LZ) on the basis of affinity. Here, we show that T cell help regulates the speed of cell cycle phase transitions and DNA replication of GC B cells. Genome sequencing and single-molecule analyses revealed that T cell help shortens S phase by regulating replication fork progression, while preserving the relative order of replication origin activation. Thus, high-affinity GC B cells are selected by a mechanism that involves prolonged dwell time in the DZ where selected cells undergo accelerated cell cycles.


Assuntos
Linfócitos B/citologia , Ciclo Celular/imunologia , Replicação do DNA/imunologia , Centro Germinativo/citologia , Imunidade Humoral/genética , Linfócitos T/imunologia , Animais , Ciclo Celular/genética , Proliferação de Células , Replicação do DNA/genética , Regulação da Expressão Gênica , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Fase S/genética , Fase S/imunologia
16.
Sci Rep ; 5: 11046, 2015 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-26078204

RESUMO

Helicobacter pylori at multiplicity of infection (MOI ≥ 50) have been shown to cause apoptosis in RAW264.7 monocytic macrophage cells. Because chronic gastric infection by H. pylori results in the persistence of macrophages in the host's gut, it is likely that H. pylori is present at low to moderate, rather than high numbers in the infected host. At present, the effect of low-MOI H. pylori infection on macrophage has not been fully elucidated. In this study, we investigated the genome-wide transcriptional regulation of H. pylori-infected RAW264.7 cells at MOI 1, 5 and 10 in the absence of cellular apoptosis. Microarray data revealed up- and down-regulation of 1341 and 1591 genes, respectively. The expression of genes encoding for DNA replication and cell cycle-associated molecules, including Aurora-B kinase (AurkB) were down-regulated. Immunoblot analysis verified the decreased expression of AurkB and downstream phosphorylation of Cdk1 caused by H. pylori infection. Consistently, we observed that H. pylori infection inhibited cell proliferation and progression through the G1/S and G2/M checkpoints. In summary, we suggest that H. pylori disrupts expression of cell cycle-associated genes, thereby impeding proliferation of RAW264.7 cells, and such disruption may be an immunoevasive strategy utilized by H. pylori.


Assuntos
Pontos de Checagem do Ciclo Celular/genética , Proteínas de Ciclo Celular/genética , Genoma , Helicobacter pylori/imunologia , Macrófagos/metabolismo , Animais , Aurora Quinase B/genética , Aurora Quinase B/imunologia , Carga Bacteriana , Proteína Quinase CDC2/genética , Proteína Quinase CDC2/imunologia , Pontos de Checagem do Ciclo Celular/imunologia , Proteínas de Ciclo Celular/imunologia , Linhagem Celular Transformada , Proliferação de Células , Replicação do DNA/genética , Replicação do DNA/imunologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Helicobacter pylori/patogenicidade , Evasão da Resposta Imune , Macrófagos/microbiologia , Camundongos , Análise de Sequência com Séries de Oligonucleotídeos , Fosforilação
17.
J Gen Virol ; 96(Pt 1): 115-117, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25304651

RESUMO

Torque teno virus (TTV) is increasingly considered a universal marker of global immune function. The virus is supposed to replicate in lymphocytes, but poor information is available about fluctuations of viraemia after administration of anti-lymphocyte agents. We studied TTV kinetics in a cohort of 70 kidney±pancreas recipients receiving one of two different anti-T-cell induction immunosuppressants. During the first 30 days after anti-T-cell antibody administration, we report kinetics of TTV viraemia compatible with replication in T lymphocytes, and highly dependent on the potency of the anti-T-cell drug administered.


Assuntos
Soro Antilinfocitário/imunologia , Replicação do DNA/genética , Imunossupressores/imunologia , Linfócitos T/imunologia , Torque teno virus/genética , Torque teno virus/imunologia , Viremia/imunologia , Replicação do DNA/imunologia , Infecções por Vírus de DNA/imunologia , Infecções por Vírus de DNA/virologia , Humanos , Cinética , Linfócitos T/virologia , Viremia/virologia
18.
J Gen Virol ; 96(Pt 1): 74-84, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25304652

RESUMO

Porcine sapovirus (PSaV) of the family Caliciviridae, is the only member of the genus Sapovirus with cell culture and reverse genetics systems. When combined with the piglet model, these approaches provide a system to understand the molecular basis of sapovirus pathogenesis. The replication of PSaV in cell culture is, however, restricted, displaying an absolute requirement for bile acids and producing lower levels of infectious virus than other caliciviruses. The effect of bile acids has previously been linked to a reduction in the signal transducer and activator of transcription (STAT1)-mediated signalling pathway. In the current study, we observed that even in the presence of bile acids, PSaV replication in cell culture was restricted by soluble factors produced from infected cells. This effect was at least partially due to secreted IFN because treatment of cells with recombinant porcine IFN-ß resulted in significantly reduced viral replication. Moreover, IFN-mediated signalling pathways (IFN, STAT1 and the 2',5'-oligoadenylate synthetase) were activated during PSaV infection. Characterization of PSaV growth in cell lines deficient in their ability to induce or respond to IFN showed a 100-150-fold increase in infectious virus production, indicating that the primary role of bile acids was not the inactivation of the innate immune response. Furthermore, the use of IFN-deficient cell lines enabled more efficient recovery of PSaV from cDNA constructs. Overall, the highly efficient cell culture and reverse genetics system established here for PSaV highlighted the key role of the innate immune response in the restriction of PSaV infection and should greatly facilitate further molecular studies on sapovirus host-cell interactions.


Assuntos
Infecções por Caliciviridae/imunologia , Replicação do DNA/genética , Interferon beta/imunologia , Sapovirus/genética , Sapovirus/imunologia , Replicação Viral/genética , Animais , Infecções por Caliciviridae/virologia , Linhagem Celular , Replicação do DNA/imunologia , DNA Complementar/genética , Imunidade Inata/imunologia , RNA Viral/genética , RNA Viral/imunologia , Suínos , Replicação Viral/imunologia
19.
PLoS One ; 9(5): e96527, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24801069

RESUMO

The heat shock protein of Toxoplasma gondii (TgHSP70) is a parasite virulence factor that is expressed during T. gondii stage conversion. To verify the effect of dexamethasone (DXM)-induced infection reactivation in the TgHSP70-specific humoral immune response and the presence of the protein in the mouse brain, we produced recombinant TgHSP70 and anti-TgHSP70 IgY antibodies to detect the protein, the specific antibody and levels of immune complexes (ICs) systemically, as well as the protein in the brain of resistant (BALB/c) and susceptible (C57BL/6) mice. It was observed higher TgHSP70-specific antibody titers in serum samples of BALB/c compared with C57BL/6 mice. However, the susceptible mice presented the highest levels of TgHSP70 systemically and no detection of specific ICs. The DXM treatment induced increased parasitism and lower inflammatory changes in the brain of C57BL/6, but did not interfere with the cerebral parasitism in BALB/c mice. Additionally, DXM treatment decreased the serological TgHSP70 concentration in both mouse lineages. C57BL/6 mice presented high expression of TgHSP70 in the brain with the progression of infection and under DXM treatment. Taken together, these data indicate that the TgHSP70 release into the bloodstream depends on the death of the parasites mediated by the host immune response, whereas the increased TgHSP70 expression in the brain depends on the multiplication rate of the parasite.


Assuntos
Encéfalo/imunologia , Replicação do DNA/imunologia , Proteínas de Choque Térmico HSP70/imunologia , Imunidade Humoral/imunologia , Parasitos/imunologia , Toxoplasma/imunologia , Toxoplasmose Animal/imunologia , Animais , Encéfalo/parasitologia , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Toxoplasmose Animal/parasitologia
20.
Clin Exp Immunol ; 175(1): 17-24, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23786362

RESUMO

Systemic autoimmunity is a complex disease process that results from a loss of immunological tolerance characterized by the inability of the immune system to discriminate self from non-self. In patients with the prototypic autoimmune disease systemic lupus erythematosus (SLE), formation of autoantibodies targeting ubiquitous nuclear antigens and subsequent deposition of immune complexes in the vascular bed induces inflammatory tissue injury that can affect virtually any organ system. Given the extraordinary genetic and phenotypic heterogeneity of SLE, one approach to the genetic dissection of complex SLE is to study monogenic diseases, for which a single gene defect is responsible. Considerable success has been achieved from the analysis of the rare monogenic disorder Aicardi-Goutières syndrome (AGS), an inflammatory encephalopathy that clinically resembles in-utero-acquired viral infection and that also shares features with SLE. Progress in understanding the cellular and molecular functions of the AGS causing genes has revealed novel pathways of the metabolism of intracellular nucleic acids, the major targets of the autoimmune attack in patients with SLE. Induction of autoimmunity initiated by immune recognition of endogenous nucleic acids originating from processes such as DNA replication/repair or endogenous retro-elements represents novel paradigms of SLE pathogenesis. These findings illustrate how investigating rare monogenic diseases can also fuel discoveries that advance our understanding of complex disease. This will not only aid the development of improved tools for SLE diagnosis and disease classification, but also the development of novel targeted therapeutic approaches.


Assuntos
Doenças Autoimunes do Sistema Nervoso , Autoimunidade/genética , Reparo do DNA , Replicação do DNA , DNA , Lúpus Eritematoso Sistêmico , Malformações do Sistema Nervoso , Animais , Autoanticorpos/genética , Autoanticorpos/imunologia , Doenças Autoimunes do Sistema Nervoso/genética , Doenças Autoimunes do Sistema Nervoso/imunologia , Doenças Autoimunes do Sistema Nervoso/patologia , Doenças Autoimunes do Sistema Nervoso/terapia , DNA/genética , DNA/imunologia , Reparo do DNA/genética , Reparo do DNA/imunologia , Replicação do DNA/genética , Replicação do DNA/imunologia , Modelos Animais de Doenças , Humanos , Lúpus Eritematoso Sistêmico/genética , Lúpus Eritematoso Sistêmico/imunologia , Lúpus Eritematoso Sistêmico/patologia , Lúpus Eritematoso Sistêmico/terapia , Malformações do Sistema Nervoso/genética , Malformações do Sistema Nervoso/imunologia , Malformações do Sistema Nervoso/patologia , Malformações do Sistema Nervoso/terapia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...