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  1 / 2328 MEDLINE  
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[PMID]:29194761
[Au] Autor:Chu FC; Klobasa W; Grubbs N; Lorenzen MD
[Ad] Endereço:Department of Entomology and Plant Pathology, North Carolina State University, Raleigh, NC, USA.
[Ti] Título:Development and use of a piggyBac-based jumpstarter system in Drosophila suzukii.
[So] Source:Arch Insect Biochem Physiol;97(3), 2018 Mar.
[Is] ISSN:1520-6327
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Spotted wing drosophila, Drosophila suzukii, is an invasive pest that primarily attacks fresh, soft-skinned fruit. Although others have reported successful integration of marked piggyBac elements into the D. suzukii genome, with a very respectable transgenesis rate of ∼16%, here we take this work a step further by creating D. suzukii jumpstarter strains. These were generated through integration of a fluorescent-marked Minos element carrying a heat shock protein 70-driven piggyBac transposase gene. We demonstrate that there is a dramatic increase in transformation rates when germline transformation is performed in a transposase-expressing background. For example, we achieved transformation rates as high as 80% when microinjecting piggyBac-based plasmids into embryos derived from one of these D. suzukii jumpstarter strains. We also investigate the effect of insert size on transformation efficiency by testing the ability of the most efficient jumpstarter strain to catalyze integration of differently-sized piggyBac elements. Finally, we demonstrate the ability of a jumpstarter strain to remobilize an already-integrated piggyBac element to a new location, demonstrating that our jumpstarter strains could be used in conjunction with a piggyBac-based donor strain for genome-wide mutagenesis of D. suzukii.
[Mh] Termos MeSH primário: Animais Geneticamente Modificados
Elementos de DNA Transponíveis
Drosophila/genética
Engenharia Genética/métodos
Mutagênese
[Mh] Termos MeSH secundário: Animais
Transposases
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Transposable Elements); EC 2.7.7.- (Transposases)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180216
[Lr] Data última revisão:
180216
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171202
[St] Status:MEDLINE
[do] DOI:10.1002/arch.21439


  2 / 2328 MEDLINE  
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[PMID]:28387938
[Au] Autor:Katayama M; Hirayama T; Tani T; Nishimori K; Onuma M; Fukuda T
[Ad] Endereço:Ecological Risk Assessment and Control Section, Center for Environmental Biology and Ecosystem, National Institute for Environmental Studies, Onogawa, Tsukuba, Ibaraki, Japan.
[Ti] Título:Chick derived induced pluripotent stem cells by the poly-cistronic transposon with enhanced transcriptional activity.
[So] Source:J Cell Physiol;233(2):990-1004, 2018 Feb.
[Is] ISSN:1097-4652
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Induced pluripotent stem (iPS) cell technology lead terminally differentiated cells into the pluripotent stem cells through the expression of defined reprogramming factors. Although, iPS cells have been established in a number of mammalian species, including mouse, human, and monkey, studies on iPS cells in avian species are still very limited. To establish chick iPS cells, six factors were used within the poly-cistronic reprogramming vector (PB-R6F), containing M3O (MyoD derived transactivation domain fused with Oct3/4), Sox2, Klf4, c-Myc, Lin28, and Nanog. The PB-R6F derived iPS cells were alkaline-phosphatase and SSEA-1 positive, which are markers of pluripotency. Elevated levels of endogenous Oct3/4 and Nanog genes were detected in the established iPS cells, suggesting the activation of the FGF signaling pathway is critical for the pluripotent status. Histological analysis of teratoma revealed that the established chick iPS cells have differentiation ability into three-germ-layer derived tissues. This is the first report of establishment of avian derived iPS cells with a single poly-cistronic transposon based expression system. The establishment of avian derived iPS cells could contribute to the genetic conservation and modification of avian species.
[Mh] Termos MeSH primário: Reprogramação Celular
Elementos de DNA Transponíveis
Fibroblastos/fisiologia
Células-Tronco Pluripotentes Induzidas/fisiologia
Transcrição Genética
Ativação Transcricional
[Mh] Termos MeSH secundário: Animais
Diferenciação Celular
Células Cultivadas
Técnicas de Reprogramação Celular
Galinhas
Técnicas de Cocultura
Feminino
Fatores de Crescimento de Fibroblastos/genética
Fatores de Crescimento de Fibroblastos/metabolismo
Fibroblastos/metabolismo
Perfilação da Expressão Gênica
Células-Tronco Pluripotentes Induzidas/metabolismo
Fenótipo
Análise de Sequência de DNA
Transdução de Sinais
Fatores de Transcrição/genética
Fatores de Transcrição/metabolismo
Transcriptoma
Transfecção
Transposases/genética
Transposases/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Transposable Elements); 0 (Transcription Factors); 62031-54-3 (Fibroblast Growth Factors); EC 2.7.7.- (Transposases)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171128
[Lr] Data última revisão:
171128
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170408
[St] Status:MEDLINE
[do] DOI:10.1002/jcp.25947


  3 / 2328 MEDLINE  
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[PMID]:28934495
[Au] Autor:Feng L; Wang G; Hamilton EP; Xiong J; Yan G; Chen K; Chen X; Dui W; Plemens A; Khadr L; Dhanekula A; Juma M; Dang HQ; Kapler GM; Orias E; Miao W; Liu Y
[Ad] Endereço:Department of Pathology, University of Michigan, Ann Arbor, MI 48109, USA.
[Ti] Título:A germline-limited piggyBac transposase gene is required for precise excision in Tetrahymena genome rearrangement.
[So] Source:Nucleic Acids Res;45(16):9481-9502, 2017 Sep 19.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Developmentally programmed genome rearrangement accompanies differentiation of the silent germline micronucleus into the transcriptionally active somatic macronucleus in the ciliated protozoan Tetrahymena thermophila. Internal eliminated sequences (IES) are excised, followed by rejoining of MAC-destined sequences, while fragmentation occurs at conserved chromosome breakage sequences, generating macronuclear chromosomes. Some macronuclear chromosomes, referred to as non-maintained chromosomes (NMC), are lost soon after differentiation. Large NMC contain genes implicated in development-specific roles. One such gene encodes the domesticated piggyBac transposase TPB6, required for heterochromatin-dependent precise excision of IES residing within exons of functionally important genes. These conserved exonic IES determine alternative transcription products in the developing macronucleus; some even contain free-standing genes. Examples of precise loss of some exonic IES in the micronucleus and retention of others in the macronucleus of related species suggest an evolutionary analogy to introns. Our results reveal that germline-limited sequences can encode genes with specific expression patterns and development-related functions, which may be a recurring theme in eukaryotic organisms experiencing programmed genome rearrangement during germline to soma differentiation.
[Mh] Termos MeSH primário: Proteínas de Protozoários/metabolismo
Tetrahymena thermophila/genética
Transposases/metabolismo
[Mh] Termos MeSH secundário: Cromossomos/genética
Éxons
Rearranjo Gênico
Heterocromatina/genética
Sequências Repetidas Invertidas
Macronúcleo/genética
Micronúcleo Germinativo
Proteínas de Protozoários/genética
Interferência de RNA
Transposases/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Heterochromatin); 0 (Protozoan Proteins); EC 2.7.7.- (Transposases)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171024
[Lr] Data última revisão:
171024
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170922
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkx652


  4 / 2328 MEDLINE  
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[PMID]:28910967
[Au] Autor:Bertels F; Gallie J; Rainey PB
[Ad] Endereço:New Zealand Institute for Advanced Study, Massey University at Albany, Auckland, New Zealand.
[Ti] Título:Identification and Characterization of Domesticated Bacterial Transposases.
[So] Source:Genome Biol Evol;9(8):2110-2121, 2017 Aug 01.
[Is] ISSN:1759-6653
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Selfish genetic elements, such as insertion sequences and transposons are found in most genomes. Transposons are usually identifiable by their high copy number within genomes. In contrast, REP-associated tyrosine transposases (RAYTs), a recently described class of bacterial transposase, are typically present at just one copy per genome. This suggests that RAYTs no longer copy themselves and thus they no longer function as a typical transposase. Motivated by this possibility we interrogated thousands of fully sequenced bacterial genomes in order to determine patterns of RAYT diversity, their distribution across chromosomes and accessory elements, and rate of duplication. RAYTs encompass exceptional diversity and are divisible into at least five distinct groups. They possess features more similar to housekeeping genes than insertion sequences, are predominantly vertically transmitted and have persisted through evolutionary time to the point where they are now found in 24% of all species for which at least one fully sequenced genome is available. Overall, the genomic distribution of RAYTs suggests that they have been coopted by host genomes to perform a function that benefits the host cell.
[Mh] Termos MeSH primário: Proteínas de Bactérias/genética
Evolução Molecular
Genoma Bacteriano
Sequências Repetitivas de Ácido Nucleico
Análise de Sequência de DNA/métodos
Transposases/classificação
Transposases/genética
[Mh] Termos MeSH secundário: Biologia Computacional/métodos
Regulação Enzimológica da Expressão Gênica
Sequências Repetidas Invertidas
Filogenia
Tirosina/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Bacterial Proteins); 42HK56048U (Tyrosine); EC 2.7.7.- (Transposases)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171107
[Lr] Data última revisão:
171107
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170916
[St] Status:MEDLINE
[do] DOI:10.1093/gbe/evx146


  5 / 2328 MEDLINE  
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[PMID]:28846090
[Au] Autor:Corces MR; Trevino AE; Hamilton EG; Greenside PG; Sinnott-Armstrong NA; Vesuna S; Satpathy AT; Rubin AJ; Montine KS; Wu B; Kathiria A; Cho SW; Mumbach MR; Carter AC; Kasowski M; Orloff LA; Risca VI; Kundaje A; Khavari PA; Montine TJ; Greenleaf WJ; Chang HY
[Ad] Endereço:Center for Personal Dynamic Regulomes, Stanford University School of Medicine, Stanford, California, USA.
[Ti] Título:An improved ATAC-seq protocol reduces background and enables interrogation of frozen tissues.
[So] Source:Nat Methods;14(10):959-962, 2017 Oct.
[Is] ISSN:1548-7105
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:We present Omni-ATAC, an improved ATAC-seq protocol for chromatin accessibility profiling that works across multiple applications with substantial improvement of signal-to-background ratio and information content. The Omni-ATAC protocol generates chromatin accessibility profiles from archival frozen tissue samples and 50-µm sections, revealing the activities of disease-associated DNA elements in distinct human brain structures. The Omni-ATAC protocol enables the interrogation of personal regulomes in tissue context and translational studies.
[Mh] Termos MeSH primário: DNA/genética
Congelamento
Genoma
Manejo de Espécimes/métodos
[Mh] Termos MeSH secundário: Animais
Encéfalo
Linhagem Celular
Eritrócitos
Regulação Enzimológica da Expressão Gênica
Estudo de Associação Genômica Ampla
Seres Humanos
Queratinócitos
Camundongos
Replicação de Sequência Autossustentável
Neoplasias da Glândula Tireoide
Transposases/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
9007-49-2 (DNA); EC 2.7.7.- (Transposases)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171010
[Lr] Data última revisão:
171010
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170829
[St] Status:MEDLINE
[do] DOI:10.1038/nmeth.4396


  6 / 2328 MEDLINE  
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[PMID]:28666380
[Au] Autor:Luo W; Galvan DL; Woodard LE; Dorset D; Levy S; Wilson MH
[Ad] Endereço:Department of Veterans Affairs, Nashville, TN 37212 USA and Department of Medicine, Department of Pharmacology, Vanderbilt University Medical Center, Nashville, TN 37232, USA.
[Ti] Título:Comparative analysis of chimeric ZFP-, TALE- and Cas9-piggyBac transposases for integration into a single locus in human cells.
[So] Source:Nucleic Acids Res;45(14):8411-8422, 2017 Aug 21.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Integrating DNA delivery systems hold promise for many applications including treatment of diseases; however, targeted integration is needed for improved safety. The piggyBac (PB) transposon system is a highly active non-viral gene delivery system capable of integrating defined DNA segments into host chromosomes without requiring homologous recombination. We systematically compared four different engineered zinc finger proteins (ZFP), four transcription activator-like effector proteins (TALE), CRISPR associated protein 9 (SpCas9) and the catalytically inactive dSpCas9 protein fused to the amino-terminus of the transposase enzyme designed to target the hypoxanthine phosphoribosyltransferase (HPRT) gene located on human chromosome X. Chimeric transposases were evaluated for expression, transposition activity, chromatin immunoprecipitation at the target loci, and targeted knockout of the HPRT gene in human cells. One ZFP-PB and one TALE-PB chimera demonstrated notable HPRT gene targeting. In contrast, Cas9/dCas9-PB chimeras did not result in gene targeting. Instead, the HPRT locus appeared to be protected from transposon integration. Supplied separately, PB permitted highly efficient isolation of Cas9-mediated knockout of HPRT, with zero transposon integrations in HPRT by deep sequencing. In summary, these tools may allow isolation of 'targeted-only' cells, be utilized to protect a genomic locus from transposon integration, and enrich for Cas9-mutated cells.
[Mh] Termos MeSH primário: Técnicas de Inativação de Genes/métodos
Marcação de Genes/métodos
Técnicas de Transferência de Genes
Mutagênese Insercional/métodos
[Mh] Termos MeSH secundário: Proteínas de Bactérias/genética
Sistemas CRISPR-Cas/genética
Linhagem Celular Tumoral
Elementos de DNA Transponíveis/genética
Endonucleases/genética
Seres Humanos
Hipoxantina Fosforribosiltransferase/genética
Hipoxantina Fosforribosiltransferase/metabolismo
Proteínas Recombinantes de Fusão/genética
Reprodutibilidade dos Testes
Nucleases dos Efetores Semelhantes a Ativadores de Transcrição/genética
Efetores Semelhantes a Ativadores de Transcrição/genética
Transposases/genética
Dedos de Zinco/genética
[Pt] Tipo de publicação:COMPARATIVE STUDY; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Bacterial Proteins); 0 (DNA Transposable Elements); 0 (Recombinant Fusion Proteins); 0 (Transcription Activator-Like Effectors); EC 2.4.2.8 (Hypoxanthine Phosphoribosyltransferase); EC 2.7.7.- (Transposases); EC 3.1.- (Cas9 endonuclease Streptococcus pyogenes); EC 3.1.- (Endonucleases); EC 3.1.- (Transcription Activator-Like Effector Nucleases)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171023
[Lr] Data última revisão:
171023
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170702
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkx572


  7 / 2328 MEDLINE  
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[PMID]:28656983
[Au] Autor:Mack SC; Suzuki H; Taylor MD
[Ad] Endereço:Cleveland Clinic, Stem Cell Biology and Regenerative Medicine, Cleveland, Ohio, USA.
[Ti] Título:Transposase-driven rearrangements in human tumors.
[So] Source:Nat Genet;49(7):975-977, 2017 Jun 28.
[Is] ISSN:1546-1718
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:A new study shows that aberrant DNA transposase activity promotes structural alterations that are clonally selected to drive tumor development. This discovery uncovers novel mechanisms of tumor-suppressor gene inactivation and highlights a new approach to cancer gene identification.
[Mh] Termos MeSH primário: Neoplasias/genética
Transposases/química
[Mh] Termos MeSH secundário: Seres Humanos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
EC 2.7.7.- (Transposases)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171026
[Lr] Data última revisão:
171026
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170629
[St] Status:MEDLINE
[do] DOI:10.1038/ng.3908


  8 / 2328 MEDLINE  
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[PMID]:28650484
[Au] Autor:Kas SM; de Ruiter JR; Schipper K; Annunziato S; Schut E; Klarenbeek S; Drenth AP; van der Burg E; Klijn C; Ten Hoeve JJ; Adams DJ; Koudijs MJ; Wesseling J; Nethe M; Wessels LFA; Jonkers J
[Ad] Endereço:Division of Molecular Pathology, The Netherlands Cancer Institute, Amsterdam, the Netherlands.
[Ti] Título:Insertional mutagenesis identifies drivers of a novel oncogenic pathway in invasive lobular breast carcinoma.
[So] Source:Nat Genet;49(8):1219-1230, 2017 Aug.
[Is] ISSN:1546-1718
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Invasive lobular carcinoma (ILC) is the second most common breast cancer subtype and accounts for 8-14% of all cases. Although the majority of human ILCs are characterized by the functional loss of E-cadherin (encoded by CDH1), inactivation of Cdh1 does not predispose mice to develop mammary tumors, implying that mutations in additional genes are required for ILC formation in mice. To identify these genes, we performed an insertional mutagenesis screen using the Sleeping Beauty transposon system in mice with mammary-specific inactivation of Cdh1. These mice developed multiple independent mammary tumors of which the majority resembled human ILC in terms of morphology and gene expression. Recurrent and mutually exclusive transposon insertions were identified in Myh9, Ppp1r12a, Ppp1r12b and Trp53bp2, whose products have been implicated in the regulation of the actin cytoskeleton. Notably, MYH9, PPP1R12B and TP53BP2 were also frequently aberrated in human ILC, highlighting these genes as drivers of a novel oncogenic pathway underlying ILC development.
[Mh] Termos MeSH primário: Neoplasias da Mama/genética
Carcinoma Lobular/genética
Mutagênese Insercional
[Mh] Termos MeSH secundário: Animais
Caderinas/genética
Linhagem Celular
Sobrevivência Celular/genética
Transformação Celular Neoplásica/genética
Feminino
Haplótipos
Seres Humanos
Masculino
Camundongos
Fosfatase de Miosina-de-Cadeia-Leve/genética
Miosina não Muscular Tipo IIA/genética
Transposases/genética
Proteínas Supressoras de Tumor/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cadherins); 0 (E-cadherin protein, mouse); 0 (Myh9 protein, mouse); 0 (Trp53bp2 protein, mouse); 0 (Tumor Suppressor Proteins); EC 2.7.7.- (Transposases); EC 3.1.3.53 (Myosin-Light-Chain Phosphatase); EC 3.1.3.53 (Ppp1r12a protein, mouse); EC 3.6.1.- (Nonmuscle Myosin Type IIA)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170921
[Lr] Data última revisão:
170921
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170627
[St] Status:MEDLINE
[do] DOI:10.1038/ng.3905


  9 / 2328 MEDLINE  
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[PMID]:28504702
[Au] Autor:Henssen AG; Koche R; Zhuang J; Jiang E; Reed C; Eisenberg A; Still E; MacArthur IC; Rodríguez-Fos E; Gonzalez S; Puiggròs M; Blackford AN; Mason CE; de Stanchina E; Gönen M; Emde AK; Shah M; Arora K; Reeves C; Socci ND; Perlman E; Antonescu CR; Roberts CWM; Steen H; Mullen E; Jackson SP; Torrents D; Weng Z; Armstrong SA; Kentsis A
[Ad] Endereço:Molecular Pharmacology Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, New York, USA.
[Ti] Título:PGBD5 promotes site-specific oncogenic mutations in human tumors.
[So] Source:Nat Genet;49(7):1005-1014, 2017 Jul.
[Is] ISSN:1546-1718
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Genomic rearrangements are a hallmark of human cancers. Here, we identify the piggyBac transposable element derived 5 (PGBD5) gene as encoding an active DNA transposase expressed in the majority of childhood solid tumors, including lethal rhabdoid tumors. Using assembly-based whole-genome DNA sequencing, we found previously undefined genomic rearrangements in human rhabdoid tumors. These rearrangements involved PGBD5-specific signal (PSS) sequences at their breakpoints and recurrently inactivated tumor-suppressor genes. PGBD5 was physically associated with genomic PSS sequences that were also sufficient to mediate PGBD5-induced DNA rearrangements in rhabdoid tumor cells. Ectopic expression of PGBD5 in primary immortalized human cells was sufficient to promote cell transformation in vivo. This activity required specific catalytic residues in the PGBD5 transposase domain as well as end-joining DNA repair and induced structural rearrangements with PSS breakpoints. These results define PGBD5 as an oncogenic mutator and provide a plausible mechanism for site-specific DNA rearrangements in childhood and adult solid tumors.
[Mh] Termos MeSH primário: Transformação Celular Neoplásica/genética
Tumor Rabdoide/genética
Transposases/fisiologia
[Mh] Termos MeSH secundário: Adulto
Animais
Domínio Catalítico
Linhagem Celular
Criança
Pré-Escolar
Aberrações Cromossômicas
Pontos de Quebra do Cromossomo
Reparo do DNA por Junção de Extremidades/genética
DNA de Neoplasias/genética
Rearranjo Gênico/genética
Genes Supressores de Tumor
Seres Humanos
Lactente
Camundongos
Camundongos Nus
Mutagênese Sítio-Dirigida
Interferência de RNA
Proteínas Recombinantes/metabolismo
Sequências Reguladoras de Ácido Nucleico
Sequências Repetidas Terminais/genética
Transposases/química
Transposases/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Neoplasm); 0 (Recombinant Proteins); EC 2.7.7.- (PGBD5 protein, human); EC 2.7.7.- (Transposases)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171115
[Lr] Data última revisão:
171115
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170516
[St] Status:MEDLINE
[do] DOI:10.1038/ng.3866


  10 / 2328 MEDLINE  
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[PMID]:28482040
[Au] Autor:Liu K; Wessler SR
[Ad] Endereço:Graduate program in Botany and Plant Sciences, University of California, Riverside, CA 92521, USA.
[Ti] Título:Transposition of Mutator-like transposable elements (MULEs) resembles hAT and Transib elements and V(D)J recombination.
[So] Source:Nucleic Acids Res;45(11):6644-6655, 2017 Jun 20.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Mutator-like transposable elements (MULEs) are widespread across fungal, plant and animal species. Despite their abundance and importance as genetic tools in plants, the transposition mechanism of the MULE superfamily was previously unknown. Discovery of the Muta1 element from Aedes aegypti and its successful transposition in yeast facilitated the characterization of key steps in Muta1 transposition. Here we show that purified transposase binds specifically to the Muta1 ends and catalyzes excision through double strand breaks (DSB) and the joining of newly excised transposon ends with target DNA. In the process, the DSB forms hairpin intermediates on the flanking DNA side. Analysis of transposase proteins containing site-directed mutations revealed the importance of the conserved DDE motif and a W residue. The transposition pathway resembles that of the V(D)J recombination reaction and the mechanism of hAT and Transib transposases including the importance of the conserved W residue in both MULEs and hATs. In addition, yeast transposition and in vitro assays demonstrated that the terminal motif and subterminal repeats of the Muta1 terminal inverted repeat also influence Muta1 transposition. Collectively, our data provides new insights to understand the evolutionary relationships between MULE, hAT and Transib elements and the V(D)J recombinase.
[Mh] Termos MeSH primário: Aedes/genética
Elementos de DNA Transponíveis
Proteínas de Insetos/genética
Transposases/genética
[Mh] Termos MeSH secundário: Aedes/enzimologia
Animais
Sequência de Bases
Domínio Catalítico
Quebras de DNA de Cadeia Dupla
Escherichia coli
Genes de Insetos
Proteínas de Insetos/química
Sequências Repetidas Invertidas
Ligação Proteica
Saccharomyces cerevisiae
Transposases/química
Recombinação V(D)J
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Transposable Elements); 0 (Insect Proteins); EC 2.7.7.- (Transposases)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171107
[Lr] Data última revisão:
171107
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170509
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkx357



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