scholarly journals NcPuf1 Is a Key Virulence Factor in Neospora caninum

Pathogens ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1019
Author(s):  
Chenrong Wang ◽  
Congshan Yang ◽  
Jing Liu ◽  
Qun Liu
Keyword(s):  
Neospora Caninum ◽  
Wide Distribution ◽  
Key Factors ◽  
Processing Bodies ◽  
Puf Proteins ◽  
P Bodies ◽  
Intracellular Parasites ◽  
Binding Factor ◽  
Puf Protein

Background: Neospora caninum is an apicomplexan parasite that infects many mammals and particularly causes abortion in cattle. The key factors in its wide distribution are its virulence and ability to transform between tachyzoite and bradyzoite forms. However, the factors are not well understood. Although Puf protein (named after Pumilio from Drosophila melanogaster and fem-3 binding factor from Caenorhabditis elegans) have a functionally conserved role in promoting proliferation and inhibiting differentiation in many eukaryotes, the function of the Puf proteins in N. caninum is poorly understood. Methods: The CRISPR/CAS9 system was used to identify and study the function of the Puf protein in N. caninum. Results: We showed that N. caninum encodes a Puf protein, which was designated NcPuf1. NcPuf1 is found in the cytoplasm in intracellular parasites and in processing bodies (P-bodies), which are reported for the first time in N. caninum in extracellular parasites. NcPuf1 is not needed for the formation of P-bodies in N. caninum. The deletion of NcPuf1 (ΔNcPuf1) does not affect the differentiation in vitro and tissue cysts formation in the mouse brain. However, ΔNcPuf1 resulted in decreases in the proliferative capacity of N. caninum in vitro and virulence in mice. Conclusions: Altogether, the disruption of NcPuf1 does not affect bradyzoites differentiation, but seriously impairs tachyzoite proliferation in vitro and virulence in mice. These results can provide a theoretical basis for the development of attenuated vaccines to prevent the infection of N. caninum.

scholarly journals Molecular cloning and characterization ofNcROP2Fam-1, a member of the ROP2 family of rhoptry proteins inNeospora caninumthat is targeted by antibodies neutralizing host cell invasionin vitro

Parasitology ◽  
2013 ◽  
Vol 140 (8) ◽  
pp. 1033-1050 ◽  
Author(s):  
FERIAL ALAEDDINE ◽  
ANDREW HEMPHILL ◽  
KARIM DEBACHE ◽  
CHRISTOPHE GUIONAUD
Keyword(s):  
Host Cell ◽  
Family Member ◽  
Vaccine Candidate ◽  
Neospora Caninum ◽  
Parasitophorous Vacuole ◽  
Host Cells ◽  
General Belief ◽  
Intracellular Parasites ◽  
Promising Vaccine Candidate

SUMMARYRecent publications demonstrated that a fragment of aNeospora caninumROP2 family member antigen represents a promising vaccine candidate. We here report on the cloning of the cDNA encoding this protein,N. caninumROP2 family member 1 (NcROP2Fam-1), its molecular characterization and localization. The protein possesses the hallmarks of ROP2 family members and is apparently devoid of catalytic activity. NcROP2Fam-1 is synthesized as a pre-pro-protein that is matured to 2 proteins of 49 and 55 kDa that localize to rhoptry bulbs. Upon invasion the protein is associated with the nascent parasitophorous vacuole membrane (PVM), evacuoles surrounding the host cell nucleus and, in some instances, the surface of intracellular parasites. Staining was also observed within the cyst wall of ‘cysts’ producedin vitro. Interestingly, NcROP2Fam-1 was also detected on the surface of extracellular parasites entering the host cells and antibodies directed against NcROP2Fam-1-specific peptides partially neutralized invasionin vitro. We conclude that, in spite of the general belief that ROP2 family proteins are intracellular antigens, NcROP2Fam-1 can also be considered as an extracellular antigen, a property that should be taken into account in further experiments employing ROP2 family proteins as vaccines.

Abstract 20439: Novel Long Intergenic Noncoding RNAs Modulate Adipose Functions in Human

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Xuan Zhang ◽  
Rachel Ballantyne ◽  
Chenyi Xue ◽  
Jane F Ferguson ◽  
Brian Gregory ◽  
...  
Keyword(s):  
Single Molecule ◽  
Noncoding Rnas ◽  
Cellular Functions ◽  
Protein Coding ◽  
Processing Bodies ◽  
P Bodies ◽  
Genome Wide ◽  
Potential Binding ◽  
Competing Endogenous Rnas

Recently long intergenic noncoding RNAs (lincRNAs) have emerged as key mediators of cellular differentiation and functions in a variety of cell systems critical to cardiovascular and metabolic disorders. To identify and investigate novel functional lincRNAs in human adipose, we performed deep high-throughput RNAseq (>200 million reads/sample) in subcutaneous adiposes of 13 health volunteers. Of an integrated dataset of 54,944 human lincRNAs, 6,558 lincRNAs were detected. Here we report 2 cytoplasmic adipose lincRNAs, linc-DMRT2 and linc-NFE2L3-1, were detected in human adipocytes but not monocytes or macrophages. Linc-DMRT2, one of the most abundant adipose lincRNAs, was markedly induced during in vitro human adipocyte differentiation. Notably, single molecule RNA FISH (fluorescence in situ hybridization) demonstrated that linc-DMRT2 were exclusively present in adipocyte cytoplasma and co-localized with processing bodies (P-bodies) marker, GW182, suggesting its potential role in modulating turnover of certain RNA species. In addition, linc-NFE2L3-1, predominantly detected in adipose and skeleton muscle, is localized near an established GWAS locus associated with waist-hip ratio adjusted BMI. We identified 4 SNPs in linc-NFE2L3-1 reaching genome wide significance for BMI (lead SNP rs10267498, P=2.73х10 -8 ). Linkage disequilibrium analysis confirmed linc-NFE2L3-1 harbors stronger GWAS signals than protein-coding genes in the locus, suggesting lincRNA might be causal for GWAS association with BMI. Bioinformatic prediction algorithms identified potential binding sites in linc-DMRT2 and linc-NFE2L3-1 for multiple microRNAs that have been demonstrated to regulate adipogenesis (e.g. miR-15 a/b and let-7) or adipocyte functions (e.g. miR-320). In summary, our data suggest that cytoplasmic linc-DMRT2 and linc-NFE2L3-1 may play important roles in adipocyte biology by functioning as competing endogenous RNAs and binding specific microRNAs that mediate adipocyte cellular functions. Genetic variation in such human linRNAs may contribute to cardiometabolic traits.

scholarly journals Functionally distinct roles for eEF2K in the control of ribosome availability and p-body abundance in sensory neurons

2021 ◽  
Author(s):  
Patrick R Smith ◽  
Sarah Loerch ◽  
Nikesh Kunder ◽  
Alexander D Stanowick ◽  
Tzu-Fang Lou ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Sensory Neurons ◽  
Molecular Mechanisms ◽  
Acceptor Site ◽  
Processing Bodies ◽  
P Bodies ◽  
Cryo Electron Microscopy ◽  
Coordinate Control ◽  
Eef2 Kinase

Processing bodies (p-bodies) are a prototypical phase-separated RNA-containing granule. Their abundance is highly dynamic and has been linked to translation. Yet, the molecular mechanisms responsible for coordinate control of the two processes are unclear. Here, we uncover key roles for eEF2 kinase (eEF2K) in the control of ribosome availability and p-body abundance. eEF2K acts on a sole known substrate, eEF2, to inhibit translation. We find that the eEF2K agonist nelfinavir abolishes p-bodies specifically in sensory neurons and impairs translation. To probe the latter, we used cryo-electron microscopy. Nelfinavir stabilizes vacant 80S ribosomes. They contain SERBP1 in place of mRNA and eEF2 in the acceptor site. Phosphorylated eEF2 associates with inactive ribosomes that resist splitting in vitro. Collectively, the data suggest that eEF2 phosphorylation defines a population of inactive ribosomes resistant to recycling and protected from degradation. Thus, eEF2K activity is central to both p-body abundance and ribosome availability in sensory neurons.

scholarly journals Dcp1-Bodies in Mouse Oocytes

2009 ◽  
Vol 20 (23) ◽  
pp. 4951-4961 ◽  
Author(s):  
Adam Swetloff ◽  
Beatrice Conne ◽  
Joachim Huarte ◽  
Jean-Luc Pitetti ◽  
Serge Nef ◽  
...  
Keyword(s):  
Translational Control ◽  
Processing Bodies ◽  
Body Protein ◽  
P Bodies ◽  
Rna Granules ◽  
Mouse Oocytes ◽  
Dead Box ◽  
Protein Partners ◽  
Protein Components

Processing bodies (P-bodies) are cytoplasmic granules involved in the storage and degradation of mRNAs. In somatic cells, their formation involves miRNA-mediated mRNA silencing. Many P-body protein components are also found in germ cell granules, such as in mammalian spermatocytes. In fully grown mammalian oocytes, where changes in gene expression depend entirely on translational control, RNA granules have not as yet been characterized. Here we show the presence of P-body-like foci in mouse oocytes, as revealed by the presence of Dcp1a and the colocalization of RNA-associated protein 55 (RAP55) and the DEAD box RNA helicase Rck/p54, two proteins associated with P-bodies and translational control. These P-body-like structures have been called Dcp1-bodies and in meiotically arrested primary oocytes, two types can be distinguished based on their size. They also have different protein partners and sensitivities to the depletion of endogenous siRNA/miRNA and translational inhibitors. However, both type progressively disappear during in vitro meiotic maturation and are virtually absent in metaphase II–arrested secondary oocytes. Moreover, this disassembly of hDcp1a-bodies is concomitant with the posttranslational modification of EGFP-hDcp1a.

scholarly journals In Vitro Induction of Neospora caninum Bradyzoites in Vero Cells Reveals Differential Antigen Expression, Localization, and Host-Cell Recognition of Tachyzoites and Bradyzoites

2004 ◽  
Vol 72 (1) ◽  
pp. 576-583 ◽  
Author(s):  
Nathalie Vonlaufen ◽  
Nicole Guetg ◽  
Arunasalam Naguleswaran ◽  
Norbert Müller ◽  
Camilla Björkman ◽  
...  
Keyword(s):  
Host Cell ◽  
Cyst Wall ◽  
Neospora Caninum ◽  
Antigen Expression ◽  
Vero Cells ◽  
Dense Granule ◽  
Host Cells ◽  
Transmission Electron ◽  
Intracellular Parasites

ABSTRACT We report on an optimized method for the in vitro culture of tissue cyst-forming Neospora caninum bradyzoites in Vero cells and the separation of viable parasites from host cells. Treatment of tachyzoite-infected Vero cell cultures with 17 μM sodium nitroprusside for 8 days severely scaled down parasite proliferation, led to reduced expression of tachyzoite surface antigens, and induced the expression of the bradyzoite marker NcBAG1 and the cyst wall antigen recognized by the monoclonal antibody MAbCC2. Transmission electron microscopy demonstrated that intracellular parasites were located within parasitophorous vacuoles that were surrounded by a cyst wall-like structure, and the dense granule antigens NcGRA1, NcGRA2, and NcGRA7 were incorporated into the cyst wall. Adhesion-invasion assays employing purified tachyzoites and bradyzoites showed that tachyzoites adhered to, and invaded, Vero cells with higher efficiency than bradyzoites. However, removal of terminal sialic acid residues from either the host cell or the parasite surface increased the invasion of Vero cells by bradyzoites, but not tachyzoites.

scholarly journals Using quantitative reconstitution to investigate multi-component condensates

RNA ◽  
2021 ◽  
pp. rna.079008.121
Author(s):  
Simon L Currie ◽  
Michael K Rosen
Keyword(s):  
Stress Granules ◽  
Lessons Learned ◽  
Sequencing Data ◽  
Processing Bodies ◽  
P Bodies ◽  
Multiple Components ◽  
In Vitro Reconstitution ◽  
Condensate Formation ◽  
Liquid Liquid Phase Separation

Many biomolecular condensates are thought to form via liquid-liquid phase separation (LLPS) of multivalent macromolecules. For those that form through this mechanism, our understanding has benefitted significantly from biochemical reconstitutions of key components and activities. Reconstitutions of RNA-based condensates to date have mostly been based on relatively simple collections of molecules. However, proteomics and sequencing data indicate that natural RNA-based condensates are enriched in hundreds to thousands of different components, and genetic data suggest multiple interactions can contribute to condensate formation to varying degrees. In this perspective we describe recent progress in understanding RNA-based condensates through different levels of biochemical reconstitutions, as a means to bridge the gap between simple in vitro reconstitution and cellular analyses. Complex reconstitutions provide insight into the formation, regulation, and functions of multi-component condensates. We focus on two RNA-protein condensate case studies: stress granules and RNA processing bodies (P bodies), and examine the evidence for cooperative interactions among multiple components promoting LLPS. An important concept emerging from these studies is that composition and stoichiometry regulate biochemical activities within condensates. Based on the lessons learned from stress granules and P bodies we discuss forward-looking approaches to understand the thermodynamic relationships between condensate components, with the goal of developing predictive models of composition and material properties, and their effects on biochemical activities. We anticipate that quantitative reconstitutions will facilitate understanding of the complex thermodynamics and functions of diverse RNA-protein condensates.

scholarly journals c-Jun N-terminal kinase phosphorylates DCP1a to control formation of P bodies

2011 ◽  
Vol 194 (4) ◽  
pp. 581-596 ◽  
Author(s):  
Katharina Rzeczkowski ◽  
Knut Beuerlein ◽  
Helmut Müller ◽  
Oliver Dittrich-Breiholz ◽  
Heike Schneider ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Processing Bodies ◽  
Messenger Ribonucleic Acid ◽  
P Bodies ◽  
Downstream Effectors ◽  
Inflammatory Stimuli ◽  
Response To Stress ◽  
Jnk Activation

Cytokines and stress-inducing stimuli signal through c-Jun N-terminal kinase (JNK) using a diverse and only partially defined set of downstream effectors. In this paper, the decapping complex subunit DCP1a was identified as a novel JNK target. JNK phosphorylated DCP1a at residue S315 in vivo and in vitro and coimmunoprecipitated and colocalized with DCP1a in processing bodies (P bodies). Sustained JNK activation by several different inducers led to DCP1a dispersion from P bodies, whereas IL-1 treatment transiently increased P body number. Inhibition of TAK1–JNK signaling also affected the number and size of P bodies and the localization of DCP1a, Xrn1, and Edc4. Transcriptome analysis further identified a central role for DCP1a in IL-1–induced messenger ribonucleic acid (mRNA) expression. Phosphomimetic mutation of S315 stabilized IL-8 but not IκBα mRNA, whereas overexpressed DCP1a blocked IL-8 transcription and suppressed p65 NF-κB nuclear activity. Collectively, these data reveal DCP1a as a multifunctional regulator of mRNA expression and suggest a novel mechanism controlling the subcellular localization of DCP1a in response to stress or inflammatory stimuli.

scholarly journals Functionally distinct roles for eEF2K in the control of ribosome availability and p-body abundance

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Patrick R. Smith ◽  
Sarah Loerch ◽  
Nikesh Kunder ◽  
Alexander D. Stanowick ◽  
Tzu-Fang Lou ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Sensory Neurons ◽  
Molecular Mechanisms ◽  
Acceptor Site ◽  
Processing Bodies ◽  
P Bodies ◽  
Cryo Electron Microscopy ◽  
Coordinate Control ◽  
Eef2 Kinase

AbstractProcessing bodies (p-bodies) are a prototypical phase-separated RNA-containing granule. Their abundance is highly dynamic and has been linked to translation. Yet, the molecular mechanisms responsible for coordinate control of the two processes are unclear. Here, we uncover key roles for eEF2 kinase (eEF2K) in the control of ribosome availability and p-body abundance. eEF2K acts on a sole known substrate, eEF2, to inhibit translation. We find that the eEF2K agonist nelfinavir abolishes p-bodies in sensory neurons and impairs translation. To probe the latter, we used cryo-electron microscopy. Nelfinavir stabilizes vacant 80S ribosomes. They contain SERBP1 in place of mRNA and eEF2 in the acceptor site. Phosphorylated eEF2 associates with inactive ribosomes that resist splitting in vitro. Collectively, the data suggest that eEF2K defines a population of inactive ribosomes resistant to recycling and protected from degradation. Thus, eEF2K activity is central to both p-body abundance and ribosome availability in sensory neurons.

RAPID IN VITRO PROPAGATION OF WESTERN GHATS CULTIVAR - COLOCASIA ESCULENTA FOR THE HIGH FREQUENCY OF PLANTLETS

2019 ◽  
Vol 7 (2) ◽  
Author(s):  
Jyothi R ◽  
Srinivasa Murthy K M ◽  
Hossein . ◽  
Veena .
Keyword(s):  
Tissue Culture ◽  
Wide Distribution ◽  
Colocasia Esculenta ◽  
Limiting Factor ◽  
Promising Alternative ◽  
Rapid Multiplication ◽  
Medical Plant ◽  
Total Fat ◽  
Planting Materials

Colocasia esculenta is commonly known as Taro, it is referred to as cocoyam in Nigeria. They are cherished for their rich taste, nutritional and medicinal properties. Every 100 g of taro corms possess 112 Kcal, 26.46 g carbohydrate, 1.50 g protein, 0.20 g total fat and 4.1g fiber (USDA National Nutrient Data Base). Besides its nutritional value, taro is used as a medical plant and provides bioactive compounds used as an anti-cancer drugs. Traditionally, cocoyams are vegetative propagated from tuber fragments, a practice that encourages pathogen distribution. Colocasia esculenta is a widely distributed food crop in the humid tropics and subtropics. Despite of its wide distribution, Taro plants are commonly infected with DsMV and other pathogens. This virus induces conspicuous mosaic, malformation, dwarfing or feathering on leaves in taro. As the results of infection, it reduces the quality and yield of taro production greatly. This virus is thus considered as a major limiting factor in the production of taro. Here plays the importance of  tissue culture plays a major role in producing the disease resistant plants round the year with high quality. For rapid multiplication and production of quality planting materials, tissue culture technology offers promising alternative compared to the traditional production methods. KEYWORDS: Colocasia esculenta, Virus, Pathogens, Conventional propagation, Micropropagation, Yield, Rapid multiplication, Quality

Electromagnetic field in Alzheimer’s disease: a literature review of recent preclinical and clinical studies

2020 ◽  
Vol 17 ◽  
Author(s):  
Reem Habib Mohamad Ali Ahmad ◽  
Marc Fakhoury ◽  
Nada Lawand
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Neurodegenerative Disorder ◽  
In Vivo Studies ◽  
Key Factors ◽  
Potential Benefits ◽  
Preclinical And Clinical Studies ◽  
The Brain

: Alzheimer’s disease (AD) is a neurodegenerative disorder characterized by the progressive loss of neurons leading to cognitive and memory decay. The main signs of AD include the irregular extracellular accumulation of amyloidbeta (Aβ) protein in the brain and the hyper-phosphorylation of tau protein inside neurons. Changes in Aβ expression or aggregation are considered key factors in the pathophysiology of sporadic and early-onset AD and correlate with the cognitive decline seen in patients with AD. Despite decades of research, current approaches in the treatment of AD are only symptomatic in nature and are not effective in slowing or reversing the course of the disease. Encouragingly, recent evidence revealed that exposure to electromagnetic fields (EMF) can delay the development of AD and improve memory. This review paper discusses findings from in vitro and in vivo studies that investigate the link between EMF and AD at the cellular and behavioural level, and highlights the potential benefits of EMF as an innovative approach for the treatment of AD.

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