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1.
Proc Natl Acad Sci U S A ; 121(21): e2401748121, 2024 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-38739789

RESUMO

Potyviridae, the largest family of plant RNA viruses, includes many important pathogens that significantly reduce the yields of many crops worldwide. In this study, we report that the 6-kilodalton peptide 1 (6K1), one of the least characterized potyviral proteins, is an endoplasmic reticulum-localized protein. AI-assisted structure modeling and biochemical assays suggest that 6K1 forms pentamers with a central hydrophobic tunnel, can increase the cell membrane permeability of Escherichia coli and Nicotiana benthamiana, and can conduct potassium in Saccharomyces cerevisiae. An infectivity assay showed that viral proliferation is inhibited by mutations that affect 6K1 multimerization. Moreover, the 6K1 or its homologous 7K proteins from other viruses of the Potyviridae family also have the ability to increase cell membrane permeability and transmembrane potassium conductance. Taken together, these data reveal that 6K1 and its homologous 7K proteins function as viroporins in viral infected cells.


Assuntos
Nicotiana , Nicotiana/virologia , Nicotiana/metabolismo , Potyviridae/genética , Potyviridae/metabolismo , Proteínas Virais/metabolismo , Proteínas Virais/genética , Permeabilidade da Membrana Celular , Retículo Endoplasmático/metabolismo , Retículo Endoplasmático/virologia , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas Viroporinas/metabolismo , Proteínas Viroporinas/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Vírus de Plantas/genética , Vírus de Plantas/fisiologia , Doenças das Plantas/virologia , Potássio/metabolismo
2.
Front Plant Sci ; 15: 1335281, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38444533

RESUMO

Introduction: Honey bee (Apis mellifera) pollination is widely used in tree fruit production systems to improve fruit set and yield. Many plant viruses can be associated with pollen or transmitted through pollination, and can be detected through bee pollination activities. Honey bees visit multiple plants and flowers in one foraging trip, essentially sampling small amounts of pollen from a wide area. Here we report metagenomics-based area-wide monitoring of plant viruses in cherry (Prunus avium) and apple (Malus domestica) orchards in Creston Valley, British Columbia, Canada, through bee-mediated pollen sampling. Methods: Plant viruses were identified in total RNA extracted from bee and pollen samples, and compared with profiles from double stranded RNA extracted from leaf and flower tissues. CVA, PDV, PNRSV, and PVF coat protein nucleotide sequences were aligned and compared for phylogenetic analysis. Results: A wide array of plant viruses were identified in both systems, with cherry virus A (CVA), prune dwarf virus (PDV), prunus necrotic ringspot virus (PNRSV), and prunus virus F (PVF) most commonly detected. Citrus concave gum associated virus and apple stem grooving virus were only identified in samples collected during apple bloom, demonstrating changing viral profiles from the same site over time. Different profiles of viruses were identified in bee and pollen samples compared to leaf and flower samples reflective of pollen transmission affinity of individual viruses. Phylogenetic and pairwise analysis of the coat protein regions of the four most commonly detected viruses showed unique patterns of nucleotide sequence diversity, which could have implications in their evolution and management approaches. Coat protein sequences of CVA and PVF were broadly diverse with multiple distinct phylogroups identified, while PNRSV and PDV were more conserved. Conclusion: The pollen virome in fruit production systems is incredibly diverse, with CVA, PDV, PNRSV, and PVF widely prevalent in this region. Bee-mediated monitoring in agricultural systems is a powerful approach to study viral diversity and can be used to guide more targeted management approaches.

3.
BMC Womens Health ; 24(1): 25, 2024 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-38184561

RESUMO

BACKGROUND: Polycystic ovary syndrome (PCOS) is a disorder characterized by hyperandrogenism, ovulatory dysfunction, and polycystic ovarian morphologic features, and PCOS is associated with infertility. PH domain Leucine-rich repeat Protein Phosphatase 1 (PHLPP1) has been shown to regulate AKT. The aim of present study is to investigate the role of PHLPP1 in PCOS. METHODS: The expression levels of PHLPP1 in dihydrotestosterone (DHT)-treated human ovarian granular KGN cells were determined by qRT-PCR and Western blot. PHLPP1 was silenced or overexpressed using lentivirus. Cell proliferation was detected by CCK-8. Apoptosis and ROS generation were analyzed by flow cytometry. Glycolysis was analyzed by measuring extracellular acidification rate (ECAR). RESULTS: DHT treatment suppressed proliferation, promoted apoptosis, enhanced ROS, and inhibited glycolysis in KGN cells. PHLPP1 silencing alleviated the DHT-induced suppression of proliferation and glycolysis, and promotion of apoptosis and ROS in KGN cells. PHLPP1 regulated cell proliferation and glycolysis in human KGN cells via the AKT signaling pathway. CONCLUSIONS: Our results showed that PHLPP1 mediates the proliferation and aerobic glycolysis activity of human ovarian granular cells through regulating AKT signaling.


Assuntos
Síndrome do Ovário Policístico , Feminino , Humanos , Proteínas Proto-Oncogênicas c-akt , Espécies Reativas de Oxigênio , Glicólise , Proteínas Nucleares , Fosfoproteínas Fosfatases/genética
4.
PLoS Pathog ; 20(1): e1011905, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38236814
5.
Methods Mol Biol ; 2724: 127-137, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-37987903

RESUMO

Infection by positive-strand RNA viruses induces extensive remodeling of the host endomembrane system in favor of viral replication and movement. The integral membrane protein 6K2 of potyviruses induces the formation of membranous virus replication vesicles at the endoplasmic reticulum exit site (ERES). The intracellular trafficking of 6K2-induced vesicles along with microfilaments requires the vesicular transport pathway, actomyosin motility system, and possibly post-Golgi compartments such as endosomes as well. Recent studies have shown that endocytosis is essential for the intracellular movement of potyviruses from the site of viral genome replication/assembly site to plasmodesmata (PD) to enter neighboring cells. In this chapter, we describe a detailed protocol of how to use endomembrane trafficking pathway-specific chemical inhibitors and organelle-selective fluorescence dye to study the trafficking of potyviral proteins and potyvirus-induced vesicles and to unravel the role of endocytosis and the endocytic pathway in potyvirus infection in Nicotiana benthamiana plants.


Assuntos
Potyvirus , Viroses , Fluorescência , Retículo Endoplasmático , Complexo de Golgi , Nicotiana , Corantes Fluorescentes
6.
Sensors (Basel) ; 23(21)2023 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-37960492

RESUMO

The hoist cage is used to lift miners in a coal mine's auxiliary shaft. Monitoring miners' unsafe behaviors and their status in the hoist cage is crucial to production safety in coal mines. In this study, a visual detection model is proposed to estimate the number and categories of miners, and to identify whether the miners are wearing helmets and whether they have fallen in the hoist cage. A dataset with eight categories of miners' statuses in hoist cages was developed for training and validating the model. Using the dataset, the classical models were trained for comparison, from which the YOLOv5s model was selected to be the basic model. Due to small-sized targets, poor lighting conditions, and coal dust and shelter, the detection accuracy of the Yolov5s model was only 89.2%. To obtain better detection accuracy, k-means++ clustering algorithm, a BiFPN-based feature fusion network, the convolutional block attention module (CBAM), and a CIoU loss function were proposed to improve the YOLOv5s model, and an attentional multi-scale cascaded feature fusion-based YOLOv5s model (AMCFF-YOLOv5s) was subsequently developed. The training results on the self-built dataset indicate that its detection accuracy increased to 97.6%. Moreover, the AMCFF-YOLOv5s model was proven to be robust to noise and light.

7.
Mol Plant ; 16(11): 1794-1810, 2023 11 06.
Artigo em Inglês | MEDLINE | ID: mdl-37777826

RESUMO

Positive-sense single-stranded RNA (+ssRNA) viruses, the most abundant viruses of eukaryotes in nature, require the synthesis of negative-sense RNA (-RNA) using their genomic (positive-sense) RNA (+RNA) as a template for replication. Based on current evidence, viral proteins are translated via viral +RNAs, whereas -RNA is considered to be a viral replication intermediate without coding capacity. Here, we report that plant and animal +ssRNA viruses contain small open reading frames (ORFs) in their -RNA (reverse ORFs [rORFs]). Using turnip mosaic virus (TuMV) as a model for plant +ssRNA viruses, we demonstrate that small proteins encoded by rORFs display specific subcellular localizations, and confirm the presence of rORF2 in infected cells through mass spectrometry analysis. The protein encoded by TuMV rORF2 forms punctuate granules that are localized in the perinuclear region and co-localized with viral replication complexes. The rORF2 protein can directly interact with the viral RNA-dependent RNA polymerase, and mutation of rORF2 completely abolishes virus infection, whereas ectopic expression of rORF2 rescues the mutant virus. Furthermore, we show that several rORFs in the -RNA of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have the ability to suppress type I interferon production and facilitate the infection of vesicular stomatitis virus. In addition, we provide evidence that TuMV might utilize internal ribosome entry sites to translate these small rORFs. Taken together, these findings indicate that the -RNA of +ssRNA viruses can also have the coding capacity and that small proteins encoded therein play critical roles in viral infection, revealing a viral proteome larger than previously thought.


Assuntos
Vírus de Plantas , Potyvirus , Viroses , Animais , RNA Viral/genética , Proteínas Virais/genética , Proteínas Virais/metabolismo , Potyvirus/genética , Vírus de Plantas/genética
8.
Entropy (Basel) ; 25(8)2023 Aug 18.
Artigo em Inglês | MEDLINE | ID: mdl-37628263

RESUMO

Rolling bearings are crucial parts of primary mine fans. In order to guarantee the safety of coal mine production, primary mine fans commonly work during regular operation and are immediately shut down for repair in case of failure. This causes the sample imbalance phenomenon in fault diagnosis (FD), i.e., there are many more normal state samples than faulty ones, seriously affecting the precision of FD. Therefore, the current study presents an FD approach for the rolling bearings of primary mine fans under sample imbalance conditions via symmetrized dot pattern (SDP) images, denoising diffusion probabilistic models (DDPMs), the image generation method, and a convolutional neural network (CNN). First, the 1D bearing vibration signal was transformed into an SDP image with significant characteristics, and the DDPM was employed to create a generated image with similar feature distributions to the real fault image of the minority class. Then, the generated images were supplemented into the imbalanced dataset for data augmentation to balance the minority class samples with the majority ones. Finally, a CNN was utilized as a fault diagnosis model to identify and detect the rolling bearings' operating conditions. In order to assess the efficiency of the presented method, experiments were performed using the regular rolling bearing dataset and primary mine fan rolling bearing data under actual operating situations. The experimental results indicate that the presented method can more efficiently fit the real image samples' feature distribution and generate image samples with higher similarity than other commonly used methods. Moreover, the diagnostic precision of the FD model can be effectively enhanced by gradually expanding and enhancing the unbalanced dataset.

9.
J Obstet Gynaecol Can ; 45(10): 102168, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37331696

RESUMO

OBJECTIVES: To prospectively assess the diagnostic accuracy of MRI and transvaginal ultrasound (TVS) as well as the prognostic value of MRI for intrauterine adhesions (IUAs), using hysteroscopy as the reference standard. DESIGN: Prospective observational study. SETTING: Tertiary medical centre. PATIENT(S): Ninety-two women with amenorrhea, hypomenorrhea, subfertility, or recurrent pregnancy loss who underwent MRI and in whom Asherman's syndrome was suspected upon TVS. INTERVENTION(S): MRI and TVS were conducted approximately 1 week before hysteroscopy. METHODS: Ninety-two patients suspected of having Asherman's syndrome were examined by MRI and TVS within 7 days of an upcoming hysteroscopy. All hysteroscopy procedures were performed during the early proliferative phase of the menstrual cycle. All hysteroscopic diagnoses were performed by an experienced expert. All MRIs were read by 2 experienced, blinded radiologists. RESULTS: MRI was highly accurate (94.57%), sensitive (98.8%), and specific (42.9%) for diagnosing IUAs with a positive predictive value of 95.5% and a negative predictive value of 75%. The diagnostic values of MRI and TVS were significantly different according to McNemar tests. Junctional zone signal and junctional zone alterations correlated with the stage of IUAs. CONCLUSION: MRI is markedly superior to TVS in terms of diagnostic accuracy for IUAs, with total agreement with hysteroscopic findings. However, the main advantage of MRI is that, unlike TVS and hysterosalpingography, it can be used to assess the risk of hysteroscopy and to predict postoperative recovery and future pregnancy based on the uterine junctional zone.


Assuntos
Ginatresia , Doenças Uterinas , Gravidez , Humanos , Feminino , Ginatresia/diagnóstico por imagem , Ginatresia/patologia , Ginatresia/cirurgia , Doenças Uterinas/diagnóstico por imagem , Útero/patologia , Histeroscopia/métodos , Aderências Teciduais/diagnóstico por imagem , Aderências Teciduais/cirurgia , Imageamento por Ressonância Magnética
10.
Nat Commun ; 14(1): 3580, 2023 06 16.
Artigo em Inglês | MEDLINE | ID: mdl-37328517

RESUMO

NONEXPRESSER OF PATHOGENESIS-RELATED GENES 1 (NPR1) is the master regulator of salicylic acid-mediated basal and systemic acquired resistance in plants. Here, we report that NPR1 plays a pivotal role in restricting compatible infection by turnip mosaic virus, a member of the largest plant RNA virus genus Potyvirus, and that such resistance is counteracted by NUCLEAR INCLUSION B (NIb), the viral RNA-dependent RNA polymerase. We demonstrate that NIb binds to the SUMO-interacting motif 3 (SIM3) of NPR1 to prevent SUMO3 interaction and sumoylation, while sumoylation of NIb by SUMO3 is not essential but can intensify the NIb-NPR1 interaction. We discover that the interaction also impedes the phosphorylation of NPR1 at Ser11/Ser15. Moreover, we show that targeting NPR1 SIM3 is a conserved ability of NIb from diverse potyviruses. These data reveal a molecular "arms race" by which potyviruses deploy NIb to suppress NPR1-mediated resistance through disrupting NPR1 sumoylation.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , RNA de Plantas/metabolismo , Sumoilação , Imunidade Vegetal/genética , Regulação da Expressão Gênica de Plantas
11.
Viruses ; 15(5)2023 05 20.
Artigo em Inglês | MEDLINE | ID: mdl-37243295

RESUMO

Healthy agroecosystems are dependent on a complex web of factors and inter-species interactions. Flowers are hubs for pathogen transmission, including the horizontal or vertical transmission of plant-viruses and the horizontal transmission of bee-viruses. Pollination by the European honey bee (Apis mellifera) is critical for industrial fruit production, but bees can also vector viruses and other pathogens between individuals. Here, we utilized commercial honey bee pollination services in blueberry (Vaccinium corymbosum) farms for a metagenomics-based bee and plant virus monitoring system. Following RNA sequencing, viruses were identified by mapping reads to a reference sequence database through the bioinformatics portal Virtool. In total, 29 unique plant viral species were found at two blueberry farms in British Columbia (BC). Nine viruses were identified at one site in Ontario (ON), five of which were not identified in BC. Ilarviruses blueberry shock virus (BlShV) and prune dwarf virus (PDV) were the most frequently detected viruses in BC but absent in ON, while nepoviruses tomato ringspot virus and tobacco ringspot virus were common in ON but absent in BC. BlShV coat protein (CP) nucleotide sequences were nearly identical in all samples, while PDV CP sequences were more diverse, suggesting multiple strains of PDV circulating at this site. Ten bee-infecting viruses were identified, with black queen cell virus frequently detected in ON and BC. Area-wide bee-mediated pathogen monitoring can provide new insights into the diversity of viruses present in, and the health of, bee-pollination ecosystems. This approach can be limited by a short sampling season, biased towards pollen-transmitted viruses, and the plant material collected by bees can be very diverse. This can obscure the origin of some viruses, but bee-mediated virus monitoring can be an effective preliminary monitoring approach.


Assuntos
Mirtilos Azuis (Planta) , Animais , Abelhas , Polinização , Ecossistema , Plantas , Pólen
12.
Mol Plant Microbe Interact ; 36(4): 199-200, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-37071003

RESUMO

Interactions between plants and microbes are ubiquitous. The outcomes of these interactions involve interkingdom communication, with myriad, diverse signals moving between microbes and their potential plant hosts. Years of biochemical, genetic, and molecular biology research have provided an overview of the landscape of the repertoires of effectors and elicitors encoded by microbes that allow them to stimulate and manipulate responses from their potential plant hosts. Similarly, considerable insight into the plant machinery and capacity for responding to microbes has been gained. The advent of new bioinformatics and modeling approaches has greatly contributed to our understanding of how these interactions occur, and it is expected that these tools, coupled with burgeoning genome sequencing data, will eventually allow the prediction of the outcome of these interactions and whether they will result in a relationship that benefits one or both partners. As a complement to these studies, cell biological studies are elucidating how cells in the plant hosts behave in response to microbial signals. Such studies have brought new attention to the indispensable role of the plant endomembrane system in determining the outcome of plant-microbe interactions. This Focus Issue addresses not only how the plant endomembrane acts locally to mediate responses to microbes but, also, the importance of the plant endomembrane beyond the plant cell borders for cross-kingdom effects. [Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 "No Rights Reserved" license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law, 2023.


Assuntos
Interações entre Hospedeiro e Microrganismos , Plantas , Plantas/microbiologia
13.
Viruses ; 15(3)2023 03 14.
Artigo em Inglês | MEDLINE | ID: mdl-36992453

RESUMO

Viruses infect all cellular life forms and cause various diseases and significant economic losses worldwide. The majority of viruses are positive-sense RNA viruses. A common feature of infection by diverse RNA viruses is to induce the formation of altered membrane structures in infected host cells. Indeed, upon entry into host cells, plant-infecting RNA viruses target preferred organelles of the cellular endomembrane system and remodel organellar membranes to form organelle-like structures for virus genome replication, termed as the viral replication organelle (VRO) or the viral replication complex (VRC). Different viruses may recruit different host factors for membrane modifications. These membrane-enclosed virus-induced replication factories provide an optimum, protective microenvironment to concentrate viral and host components for robust viral replication. Although different viruses prefer specific organelles to build VROs, at least some of them have the ability to exploit alternative organellar membranes for replication. Besides being responsible for viral replication, VROs of some viruses can be mobile to reach plasmodesmata (PD) via the endomembrane system, as well as the cytoskeleton machinery. Viral movement protein (MP) and/or MP-associated viral movement complexes also exploit the endomembrane-cytoskeleton network for trafficking to PD where progeny viruses pass through the cell-wall barrier to enter neighboring cells.


Assuntos
Vírus de Plantas , Vírus de RNA , RNA , Replicação Viral , Citoesqueleto/metabolismo , Replicação do RNA , Vírus de RNA/genética , Proteínas Virais/genética , Vírus de Plantas/genética
14.
Sci China Life Sci ; 66(7): 1665-1681, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-36917406

RESUMO

Multiple viral infections in insect vectors with synergistic effects are common in nature, but the underlying mechanism remains elusive. Here, we find that rice gall dwarf reovirus (RGDV) facilitates the transmission of rice stripe mosaic rhabdovirus (RSMV) by co-infected leafhopper vectors. RSMV nucleoprotein (N) alone activates complete anti-viral autophagy, while RGDV nonstructural protein Pns11 alone induces pro-viral incomplete autophagy. In co-infected vectors, RSMV exploits Pns11-induced autophagosomes to assemble enveloped virions via N-Pns11-ATG5 interaction. Furthermore, RSMV could effectively propagate in Sf9 cells. Expression of Pns11 in Sf9 cells or leafhopper vectors causes the recruitment of N from the ER to Pns11-induced autophagosomes and inhibits N-induced complete autophagic flux, finally facilitating RSMV propagation. In summary, these results demonstrate a previously unappreciated role of autophagy in the regulation of the direct synergistic interaction during co-transmission of two distinct arboviruses by insect vectors and reveal the functional importance of virus-induced autophagosomes in rhabdovirus assembly.


Assuntos
Arbovírus , Hemípteros , Oryza , Reoviridae , Animais , Replicação Viral , Proteínas não Estruturais Virais/metabolismo , Hemípteros/metabolismo , Reoviridae/metabolismo , Autofagia , Insetos Vetores , Oryza/metabolismo
15.
Mol Plant ; 16(3): 632-642, 2023 03 06.
Artigo em Inglês | MEDLINE | ID: mdl-36597359

RESUMO

RNA quality control nonsense-mediated decay is involved in viral restriction in both plants and animals. However, it is not known whether two other RNA quality control pathways, nonstop decay and no-go decay, are capable of restricting viruses in plants. Here, we show that the evolutionarily conserved Pelota-Hbs1 complex negatively regulates infection of plant viruses in the family Potyviridae (termed potyvirids), the largest group of plant RNA viruses that accounts for more than half of the viral crop damage worldwide. Pelota enables the recognition of the functional G1-2A6-7 motif in the P3 cistron, which is conserved in almost all potyvirids. This allows Pelota to target the virus and act as a viral restriction factor. Furthermore, Pelota interacts with the SUMO E2-conjugating enzyme SCE1 and is SUMOylated in planta. Blocking Pelota SUMOylation disrupts the ability to recruit Hbs1 and inhibits viral RNA degradation. These findings reveal the functional importance of Pelota SUMOylation during the infection of potyvirids in plants.


Assuntos
Degradação do RNAm Mediada por Códon sem Sentido , Sumoilação , RNA , Plantas , Potyvirus
16.
BMC Plant Biol ; 23(1): 56, 2023 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-36698067

RESUMO

BACKGROUND: Areca palm (Areca catechu) is a woody perennial plant of both economical and medicinal importance grown in tropical and subtropical climates. Yet, the molecular biology study of areca palm is extremely impeded by its unavailability of a transformation method. An efficient protoplast isolation and transformation system could be highly desirable to overcome this barrier. RESULTS: Here, we described a simple and efficient method for protoplast isolation and transformation from the perennial plant areca palm. A high yield of protoplasts (2.5 × 107 protoplasts per gram of fresh leaf tissues) was obtained from the fresh light green leaflet from the newly-emerged leaf digested overnight in the enzyme solution [2% (w/v) cellulase R10, 0.5% (w/v) macerozyme R10, 0.7 M mannitol, 10 mM CaCl2, 20 mM KCl, 20 mM MES and 0.1% (w/v) BSA, pH 5.7] by the direct leaf-peeling method. The isolated areca protoplasts maintain viability of 86.6% and have been successfully transformed with a green fluorescent protein (GFP)-tagged plasmid (pGreen0029-GFP, 6.0 kb) via the polyethylene glycol (PEG)-mediated transformation. Moreover, the mannitol concentration (optimal: 0.7 M) was determined as a key factor affecting areca protoplast isolation. We also demonstrated that the optimal density of areca protoplast for efficient transformation was at 1.0-1.5 × 106 cells/ml. With the optimization of transformation parameters, we have achieved a relatively high transformation efficiency of nearly 50%. CONCLUSION: We have established the first efficient protocol for the high-yield isolation and transformation of areca palm protoplasts. This method shall be applied in various biological studies of areca palm, such as gene function analysis, genome editing, protein trafficking and localization and protein-protein interaction. In addition, the protoplast system offers a great genetic transformation approach for the woody perennial plant-areca palm. Moreover, the established platform may be applied in protoplast isolation and transformation for other important species in the palm family, including oil palm and coconut.


Assuntos
Areca , Arecaceae , Protoplastos/metabolismo , Folhas de Planta
17.
Autophagy ; 19(2): 616-631, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-35722949

RESUMO

Mitophagy that selectively eliminates damaged mitochondria is an essential mitochondrial quality control mechanism. Recently, mitophagy has been shown to be induced in host cells infected by a few animal viruses. Here, we report that southern rice black-streaked dwarf virus (SRBSDV), a plant nonenveloped double-stranded RNA virus, can also trigger mitophagy in its planthopper vector to prevent mitochondria-dependent apoptosis and promote persistent viral propagation. We find that the fibrillar structures constructed by the nonstructural protein P7-1 of SRBSDV directly target mitochondria via interaction with the mitophagy receptor BNIP3 (BCL2 interacting protein 3), and these mitochondria are then sequestered within autophagosomes to form mitophagosomes. Moreover, SRBSDV infection or P7-1 expression alone can promote BNIP3 dimerization on the mitochondria, and induce autophagy via the P7-1-ATG8 interaction. Furthermore, SRBSDV infection stimulates the phosphorylation of AMP-activated protein kinase (AMPK), resulting in BNIP3 phosphorylation via the AMPKα-BNIP3 interaction. Together, P7-1 induces BNIP3-mediated mitophagy by promoting the formation of phosphorylated BNIP3 dimers on the mitochondria. Silencing of ATG8, BNIP3, or AMPKα significantly reduces virus-induced mitophagy and viral propagation in insect vectors. These data suggest that in planthopper, SRBSDV-induced mitophagosomes are modified to accommodate virions and facilitate persistent viral propagation. In summary, our results demonstrate a previously unappreciated role of a viral protein in the induction of BNIP3-mediated mitophagy by bridging autophagosomes and mitochondria and reveal the functional importance of virus-induced mitophagy in maintaining persistent viral infection in insect vectors.Abbreviations: AMPK: AMP-activated protein kinase; ATG: autophagy related; BNIP3: BCL2 interacting protein 3; CASP3: caspase 3; dsRNA: double strand RNA; ER: endoplasmic reticulum; FITC: fluorescein isothiocyanate; FKBP8: FKBP prolyl isomerase 8; FUNDC1: FUN14 domain containing 1; GFP: green fluorescent protein; GST: glutathione S-transferase; padp: post-first access to diseased plants; Phos-tag: Phosphate-binding tag; PINK1: PTEN induced kinase 1; Sf9: Spodoptera frugiperda; SQSTM1: sequestosome 1; SRBSDV: southern rice black-streaked dwarf virus; STK11/LKB1: serine/threonine kinase 11; TOMM20: translocase of outer mitochondrial membrane 20; RBSDV: rice black-streaked dwarf virus; TUNEL: terminal deoxynucleotidyl dUTP nick end labeling; ULK1: unc-51 like autophagy activating kinase 1; VDAC1: voltage dependent anion channel 1.


Assuntos
Proteínas Quinases Ativadas por AMP , Mitofagia , Animais , Proteínas Quinases Ativadas por AMP/genética , Autofagia , Insetos Vetores , Mitofagia/genética , Infecção Persistente , Proteínas Proto-Oncogênicas c-bcl-2/genética , RNA de Cadeia Dupla , Proteínas de Membrana/metabolismo
18.
Sensors (Basel) ; 22(24)2022 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-36560323

RESUMO

Rolling bearings are a vital component of mechanical equipment. It is crucial to implement rolling bearing fault diagnosis research to guarantee the stability of the long-term action of mechanical equipment. Conversion of rolling bearing vibration signals into images for fault diagnosis research has been a practical diagnostic approach. The current paper presents a rolling bearing fault diagnosis method using symmetrized dot pattern (SDP) images and a deep residual network with convolutional block attention module (CBAM-DRN). The rolling bearing vibration signal is first visualized and transformed into an SDP image with distinct fault characteristics. Then, CBAM-DRN is utilized to derive characteristics directly and detect faults from the input SDP images. In order to prevent conventional time-frequency images from being limited by their inherent flaws and avoid missing the fault features, the SDP technique is employed to convert vibration signals into images for visualization. DRN enables adequate extraction of rolling bearing fault characteristics and prevents training difficulties and gradient vanishing in deep level networks. CBAM assists the diagnostic model in concentrating on the image's more distinctive parts and preventing the interference of non-featured parts. Finally, the method's validity was tested with a composite fault dataset of motor bearings containing multiple loads and fault diameters. The experimental results reflect that the presented approach can attain a diagnostic precision of over 99% and good stability and generalization.


Assuntos
Inteligência , Registros , Vibração
19.
Viruses ; 14(6)2022 05 24.
Artigo em Inglês | MEDLINE | ID: mdl-35746594

RESUMO

This review summarizes the history and current state of the known genetic basis for soybean resistance to Soybean mosaic virus (SMV), and examines how the integration of molecular markers has been utilized in breeding for crop improvement. SVM causes yield loss and seed quality reduction in soybean based on the SMV strain and the host genotype. Understanding the molecular underpinnings of SMV-soybean interactions and the genes conferring resistance to SMV has been a focus of intense research interest for decades. Soybean reactions are classified into three main responses: resistant, necrotic, or susceptible. Significant progress has been achieved that has greatly increased the understanding of soybean germplasm diversity, differential reactions to SMV strains, genotype-strain interactions, genes/alleles conferring specific reactions, and interactions among resistance genes and alleles. Many studies that aimed to uncover the physical position of resistance genes have been published in recent decades, collectively proposing different candidate genes. The studies on SMV resistance loci revealed that the resistance genes are mainly distributed on three chromosomes. Resistance has been pyramided in various combinations for durable resistance to SMV strains. The causative genes are still elusive despite early successes in identifying resistance alleles in soybean; however, a gene at the Rsv4 locus has been well validated.


Assuntos
Glycine max , Potyvirus , Genes de Plantas , Pesquisa em Genética , Melhoramento Vegetal , Doenças das Plantas/genética , Potyvirus/genética , Glycine max/genética
20.
PLoS Pathog ; 18(5): e1010506, 2022 05.
Artigo em Inglês | MEDLINE | ID: mdl-35533206

RESUMO

Viruses can hijack autophagosomes as the nonlytic release vehicles in cultured host cells. However, how autophagosome-mediated viral spread occurs in infected host tissues or organs in vivo remains poorly understood. Here, we report that an important rice reovirus, rice gall dwarf virus (RGDV) hijacks autophagosomes to traverse multiple insect membrane barriers in the midgut and salivary gland of leafhopper vector to enhance viral spread. Such virus-containing double-membraned autophagosomes are prevented from degradation, resulting in increased viral propagation. Mechanistically, viral nonstructural protein Pns11 induces autophagy and embeds itself in the autophagosome membranes. The autophagy-related protein 5 (ATG5)-ATG12 conjugation is essential for initial autophagosome membrane biogenesis. RGDV Pns11 specifically interacts with ATG5, both in vitro and in vivo. Silencing of ATG5 or Pns11 expression suppresses ATG8 lipidation, autophagosome formation, and efficient viral propagation. Thus, Pns11 could directly recruit ATG5-ATG12 conjugation to induce the formation of autophagosomes, facilitating viral spread within the insect bodies. Furthermore, Pns11 potentially blocks autophagosome degradation by directly targeting and mediating the reduced expression of N-glycosylated Lamp1 on lysosomal membranes. Taken together, these results highlight how RGDV remodels autophagosomes to benefit viral propagation in its insect vector.


Assuntos
Orthoreovirus , Oryza , Reoviridae , Animais , Autofagossomos/metabolismo , Autofagia , Insetos Vetores , Insetos/metabolismo , Oryza/metabolismo , Reoviridae/metabolismo , Proteínas não Estruturais Virais/genética , Proteínas não Estruturais Virais/metabolismo , Replicação Viral
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