Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 10 de 10
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Fr J Urol ; 34(4): 102590, 2024 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-38394985

RESUMO

INTRODUCTION: The objective of this study is to make an inventory of surgical practices and their consequences in the short and medium term on sexuality and micturition comfort. MATERIALS AND METHODS: It is a retrospective multicenter study over ten years on 63 men who had an operation for a fracture of the corpora cavernosa associated or not with a urethral lesion. Patient history, clinical presentation, surgical management as well as postoperative data were collected from operative reports. Residual penis curvature, IIEF5 score, IPSS score and residual pain were collected during a telephone interview during data collection. RESULTS: No statistically significant difference was demonstrated for IIEF5, IPSS, sequelae curvature, pain during intercourse, time to resumption of sexual life, rate of surgical resumption between use of absorbable or non-absorbable threads and between the realization of an overlock or a separate point. We found a significant difference in the time taken to resume sexual activity, between surgical exploration by degloving compared to elective surgical exploration. CONCLUSION: Our study shows great variability in the surgical management of penile fractures, with no influence in the short and medium term on sexuality and urination comfort. Medium-term complications such as erectile dysfunction, curvature of the penis and pain during sexual intercourse seem frequent and insufficiently diagnosed, but decrease during urological follow-up.

2.
Methods Mol Biol ; 2209: 363-385, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33201481

RESUMO

RTL (RNase three-like) proteins belong to a distinct family of endonucleases that cleave double-stranded RNAs in plants. RTL1 to 3 are structurally related to the RNAse III from E. coli and formally belong to the class 1 of RNase III proteins. RTLs have conserved RNase III signature motif(s) and up to two dsRNA binding (DRB) domains. RTLs target and cleave coding and noncoding dsRNAs, including precursors of ribosomal (rRNA), small interference (siRNA), and micro (miRNA) RNAs. Interestingly, RTL proteins have stronger affinity than RNase III-Dicer proteins for dsRNA precursors of siRNAs, but not for miRNAs. However, very little is known of the structural and molecular bases directing and controlling RTL-RNA binding and activity. To address these questions, we have developed in vitro cleavage assays that combine recombinant RTL1 protein and in vitro transcribed or plant-extracted RNAs, RT-PCR, and primer extension experiments or analysis.


Assuntos
Proteínas de Arabidopsis/metabolismo , Ensaios Enzimáticos/métodos , RNA de Plantas/metabolismo , Proteínas Repressoras/metabolismo , Arabidopsis/metabolismo , Ligação Proteica , Proteínas Recombinantes/metabolismo
3.
Plant Physiol ; 177(1): 311-327, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29622687

RESUMO

Despite intensive searches, few proteins involved in telomere homeostasis have been identified in plants. Here, we used pull-down assays to identify potential telomeric interactors in the model plant species Arabidopsis (Arabidopsis thaliana). We identified the candidate protein GH1-HMGA1 (also known as HON4), an uncharacterized linker histone protein of the High Mobility Group Protein A (HMGA) family in plants. HMGAs are architectural transcription factors and have been suggested to function in DNA damage repair, but their precise biological roles remain unclear. Here, we show that GH1-HMGA1 is required for efficient DNA damage repair and telomere integrity in Arabidopsis. GH1-HMGA1 mutants exhibit developmental and growth defects, accompanied by ploidy defects, increased telomere dysfunction-induced foci, mitotic anaphase bridges, and degraded telomeres. Furthermore, mutants have a higher sensitivity to genotoxic agents such as mitomycin C and γ-irradiation. Our work also suggests that GH1-HMGA1 is involved directly in the repair process by allowing the completion of homologous recombination.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Dano ao DNA , Reparo do DNA , Proteínas HMGA/metabolismo , Histonas/metabolismo , Telômero/metabolismo , Arabidopsis/crescimento & desenvolvimento , Cromatina/metabolismo , DNA Bacteriano/genética , Fluorescência , Recombinação Homóloga/genética , Mutação/genética , Ligação Proteica , Telomerase/metabolismo , Proteínas de Ligação a Telômeros/metabolismo
4.
Nucleic Acids Res ; 46(5): 2432-2445, 2018 03 16.
Artigo em Inglês | MEDLINE | ID: mdl-29346668

RESUMO

Replicative erosion of telomeres is naturally compensated by telomerase and studies in yeast and vertebrates show that homologous recombination can compensate for the absence of telomerase. We show that RAD51 protein, which catalyzes the key strand-invasion step of homologous recombination, is localized at Arabidopsis telomeres in absence of telomerase. Blocking the strand-transfer activity of the RAD51 in telomerase mutant plants results in a strikingly earlier onset of developmental defects, accompanied by increased numbers of end-to-end chromosome fusions. Imposing replication stress through knockout of RNaseH2 increases numbers of chromosome fusions and reduces the survival of these plants deficient for telomerase and homologous recombination. This finding suggests that RAD51-dependent homologous recombination acts as an essential backup to the telomerase for compensation of replicative telomere loss to ensure genome stability. Furthermore, we show that this positive role of RAD51 in telomere stability is dependent on the RTEL1 helicase. We propose that a RAD51 dependent break-induced replication process is activated in cells lacking telomerase activity, with RTEL1 responsible for D-loop dissolution after telomere replication.


Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/genética , DNA Helicases/fisiologia , Rad51 Recombinase/fisiologia , Encurtamento do Telômero , Arabidopsis/enzimologia , Proteínas de Arabidopsis/análise , Replicação do DNA , Instabilidade Genômica , Recombinação Homóloga , Mutação , Rad51 Recombinase/análise , Sequências Repetitivas de Ácido Nucleico , Ribonucleases/genética , Processos Estocásticos , Telomerase/genética , Telômero/química
5.
Nucleic Acids Res ; 45(20): 11891-11907, 2017 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-28981840

RESUMO

RNase III enzymes cleave double stranded (ds)RNA. This is an essential step for regulating the processing of mRNA, rRNA, snoRNA and other small RNAs, including siRNA and miRNA. Arabidopsis thaliana encodes nine RNase III: four DICER-LIKE (DCL) and five RNASE THREE LIKE (RTL). To better understand the molecular functions of RNase III in plants we developed a biochemical assay using RTL1 as a model. We show that RTL1 does not degrade dsRNA randomly, but recognizes specific duplex sequences to direct accurate cleavage. Furthermore, we demonstrate that RNase III and dsRNA binding domains (dsRBD) are both required for dsRNA cleavage. Interestingly, the four DCL and the three RTL that carry dsRBD share a conserved cysteine (C230 in Arabidopsis RTL1) in their dsRBD. C230 is essential for RTL1 and DCL1 activities and is subjected to post-transcriptional modification. Indeed, under oxidizing conditions, glutathionylation of C230 inhibits RTL1 cleavage activity in a reversible manner involving glutaredoxins. We conclude that the redox state of the dsRBD ensures a fine-tune regulation of dsRNA processing by plant RNase III.


Assuntos
Proteínas de Arabidopsis/metabolismo , Cisteína/metabolismo , RNA de Cadeia Dupla/metabolismo , RNA de Plantas/metabolismo , Proteínas Repressoras/metabolismo , Regiões 3' não Traduzidas/genética , Sequência de Aminoácidos , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Sequência de Bases , Cisteína/genética , Glutationa/metabolismo , Modelos Moleculares , Conformação de Ácido Nucleico , Oxirredução , Domínios Proteicos , Clivagem do RNA , RNA de Cadeia Dupla/química , RNA de Cadeia Dupla/genética , RNA de Plantas/química , RNA de Plantas/genética , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Motivos de Ligação ao RNA/genética , Proteínas Repressoras/química , Proteínas Repressoras/genética , Ribonuclease III/genética , Ribonuclease III/metabolismo , Homologia de Sequência do Ácido Nucleico
6.
PLoS Biol ; 13(12): e1002326, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26696443

RESUMO

Small RNAs play essential regulatory roles in genome stability, development, and responses to biotic and abiotic stresses in most eukaryotes. In plants, the RNaseIII enzyme DICER-LIKE1 (DCL1) produces miRNAs, whereas DCL2, DCL3, and DCL4 produce various size classes of siRNAs. Plants also encode RNASE THREE-LIKE (RTL) enzymes that lack DCL-specific domains and whose function is largely unknown. We found that virus infection induces RTL1 expression, suggesting that this enzyme could play a role in plant-virus interaction. To first investigate the biochemical activity of RTL1 independent of virus infection, small RNAs were sequenced from transgenic plants constitutively expressing RTL1. These plants lacked almost all DCL2-, DCL3-, and DCL4-dependent small RNAs, indicating that RTL1 is a general suppressor of plant siRNA pathways. In vivo and in vitro assays revealed that RTL1 prevents siRNA production by cleaving dsRNA prior to DCL2-, DCL3-, and DCL4-processing. The substrate of RTL1 cleavage is likely long-perfect (or near-perfect) dsRNA, consistent with the RTL1-insensitivity of miRNAs, which derive from DCL1-processing of short-imperfect dsRNA. Virus infection induces RTL1 mRNA accumulation, but viral proteins that suppress RNA silencing inhibit RTL1 activity, suggesting that RTL1 has evolved as an inducible antiviral defense that could target dsRNA intermediates of viral replication, but that a broad range of viruses counteract RTL1 using the same protein toolbox used to inhibit antiviral RNA silencing. Together, these results reveal yet another level of complexity in the evolutionary battle between viruses and plant defenses.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/virologia , Regulação da Expressão Gênica de Plantas , Interações Hospedeiro-Patógeno , Vírus de RNA/fisiologia , RNA de Plantas/antagonistas & inibidores , RNA Interferente Pequeno/antagonistas & inibidores , Proteínas Repressoras/metabolismo , Substituição de Aminoácidos , Arabidopsis/enzimologia , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Carmovirus/fisiologia , Biologia Computacional/métodos , Cucumovirus/fisiologia , Isoenzimas/genética , Isoenzimas/metabolismo , Mutagênese Sítio-Dirigida , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/virologia , Mutação Puntual , RNA Mensageiro/metabolismo , RNA de Plantas/metabolismo , RNA Interferente Pequeno/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Repressoras/genética , Tobamovirus/fisiologia , Tymovirus/fisiologia
7.
DNA Repair (Amst) ; 10(6): 611-9, 2011 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-21530420

RESUMO

Double-strand breaks in genomic DNA (DSB) are potentially lethal lesions which separate parts of chromosome arms from their centromeres. Repair of DSB by recombination can generate mutations and further chromosomal rearrangements, making the regulation of recombination and the choice of recombination pathways of the highest importance. Although knowledge of recombination mechanisms has considerably advanced, the complex interrelationships and regulation of pathways are far from being fully understood. We analyse the different pathways of DSB repair acting in G2/M phase nuclei of irradiated plants, through quantitation of the kinetics of appearance and loss of γ-H2AX foci in Arabidopsis mutants. These analyses show the roles for the four major recombination pathways in post-S-phase DSB repair and that non-homologous recombination pathways constitute the major response. The data suggest a hierarchical organisation of DSB repair in these cells: C-NHEJ acts prior to B-NHEJ which can also inhibit MMEJ. Surprisingly the quadruple ku80 xrcc1 xrcc2 xpf mutant can repair DSB, although with severely altered kinetics. This repair leads to massive genetic instability with more than 50% of mitoses showing anaphase bridges following irradiation. This study thus clarifies the relationships between the different pathways of DSB repair in the living plant and points to the existence of novel DSB repair processes.


Assuntos
Arabidopsis/metabolismo , Reparo do DNA , DNA de Plantas/metabolismo , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/efeitos da radiação , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Quebras de DNA de Cadeia Dupla , DNA de Plantas/genética , Cinética
9.
Plant J ; 64(2): 280-90, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21070408

RESUMO

Double-strand breakage (DSB) of DNA involves loss of information on the two strands of the DNA fibre and thus cannot be repaired by simple copying of the complementary strand which is possible with single-strand DNA damage. Homologous recombination (HR) can precisely repair DSB using another copy of the genome as template and non-homologous recombination (NHR) permits repair of DSB with little or no dependence on DNA sequence homology. In addition to the well-characterised Ku-dependent non-homologous end-joining (NHEJ) pathway, much recent attention has been focused on Ku-independent NHR. The complex interrelationships and regulation of NHR pathways remain poorly understood, even more so in the case of plants, and we present here an analysis of Ku-dependent and Ku-independent repair of DSB in Arabidopsis thaliana. We have characterised an Arabidopsis xrcc1 mutant and developed quantitative analysis of the kinetics of appearance and loss of γ-H2AX foci as a tool to measure DSB repair in dividing root tip cells of γ-irradiated plants in vivo. This approach has permitted determination of DSB repair kinetics in planta following a short pulse of γ-irradiation, establishing the existence of a Ku-independent, Xrcc1-dependent DSB repair pathway. Furthermore, our data show a role for Ku80 during the first minutes post-irradiation and that Xrcc1 also plays such a role, but only in the absence of Ku. The importance of Xrcc1 is, however, clearly visible at later times in the presence of Ku, showing that alternative end-joining plays an important role in DSB repair even in the presence of active NHEJ.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Quebras de DNA de Cadeia Dupla , DNA Helicases/metabolismo , Reparo do DNA , Proteínas de Ligação a DNA/metabolismo , Arabidopsis/genética , Proteínas de Ligação a DNA/genética , Raios gama , Cinética , Meristema/metabolismo , Mutação , Proteína 1 Complementadora Cruzada de Reparo de Raio-X
10.
Plant Cell ; 22(9): 3020-33, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20876831

RESUMO

Signaling of chromosomal DNA breaks is of primary importance for initiation of repair and, thus, for global genomic stability. Although the Mre11-Rad50-Nbs1 (MRN) complex is the first sensor of double-strand breaks, its role in double-strand break (DSB) signaling is not fully understood. We report the absence of γ-ray-induced, ATM/ATR-dependent histone H2AX phosphorylation in Arabidopsis thaliana rad50 and mre11 mutants, confirming that the MRN complex is required for H2AX phosphorylation by the ATM and ATR kinases in response to irradiation-induced DSB in Arabidopsis. rad50 and mre11 mutants spontaneously activate a DNA damage response, as shown by the presence of γ-H2AX foci and activation of cell cycle arrest in nonirradiated plants. This response is ATR dependent as shown both by the absence of these spontaneous foci and by the wild-type mitotic indices of double rad50 atr and mre11 atr plants. EdU S-phase labeling and fluorescence in situ hybridization analysis using specific subtelomeric probes point to a replicative S-phase origin of this chromosome damage in the double mutants and not to telomere destabilization. Thus, the data presented here show the exclusive involvement of ATR in DNA damage signaling in MRN mutants and provide evidence for a role for ATR in the avoidance of S-phase DNA damage.


Assuntos
Arabidopsis/enzimologia , Instabilidade Cromossômica , Dano ao DNA , Histonas/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Arabidopsis/genética , Arabidopsis/efeitos da radiação , Proteínas de Arabidopsis/metabolismo , Proteínas de Ligação a DNA/metabolismo , Raios gama , Proteína Homóloga a MRE11 , Mutagênese Insercional , Fosforilação
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...