scholarly journals Interaction between SWP9 and Polar Tube Proteins of the Microsporidian Nosema bombycis and Function of SWP9 as a Scaffolding Protein Contribute to Polar Tube Tethering to the Spore Wall

2016 ◽  
Vol 85 (3) ◽  
Author(s):  
Donglin Yang ◽  
Lixia Pan ◽  
Pai Peng ◽  
Xiaoqun Dang ◽  
Chunfeng Li ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Spore Wall ◽  
Scaffolding Protein ◽  
Dependent Manner ◽  
Polar Tube ◽  
Content Type ◽  
Nosema Bombycis ◽  
Invasion Mechanism ◽  
In The Beginning ◽  
And Function

ABSTRACTAll microsporidia possess a unique, highly specialized invasion mechanism that involves the polar tube and spore wall. The interaction between spore wall proteins (SWPs) and polar tube proteins (PTPs) in the formation, arrangement, orderly orientation, and function of the polar tube and spore wall remains to be determined. This study was undertaken to examine the protein interactions ofNosema bombycisSWP7 (NbSWP7), NbSWP9, and PTPs. Coimmunoprecipitation, liquid chromatography-tandem mass spectrometry (LC-MS/MS), and yeast two-hybrid data demonstrated that NbSWP9, but not NbSWP7, interacts with NbPTP1 and NbPTP2. Furthermore, immunoelectron microscopy (IEM) showed that NbSWP9 was localized mainly in the developing polar tube of sporoblasts, while NbSWP7 was found randomly in the cytoplasm. However, both NbSWP9 and NbSWP7 were located in the polar tube and spore wall ofN. bombycismature spores. The reason why NbSWP7 was localized to the polar tube may be due to the interaction between NbSWP9 and NbSWP7. Interestingly, the majority of NbSWP9, but not NbSWP7, accumulated in the beginning part of the extruded polar tube and the ruptured spore wall called the anchoring disk (AD) when the mature spores germinated under weak-alkaline environmental stimulation. Additionally, anti-NbSWP9 antibody reduced spore germination in a dose-dependent manner. In conclusion, our study further confirmed that NbSWP9 is a scaffolding protein that not only anchors and holds the polar tube but also tethers the polar tube to the spore wall.

scholarly journals Interaction and Assembly of Two Novel Proteins in the Spore Wall of the Microsporidian Species Nosema bombycis and Their Roles in Adherence to and Infection of Host Cells

2015 ◽  
Vol 83 (4) ◽  
pp. 1715-1731 ◽  
Author(s):  
Donglin Yang ◽  
Guoqing Pan ◽  
Xiaoqun Dang ◽  
Yawei Shi ◽  
Chunfeng Li ◽  
...  
Keyword(s):  
Host Cell ◽  
Spore Wall ◽  
Host Cells ◽  
Scaffolding Protein ◽  
Environmental Pressures ◽  
Content Type ◽  
Nosema Bombycis ◽  
Major Function ◽  
Microsporidian Species

Microsporidia are obligate intracellular parasites with rigid spore walls that protect against various environmental pressures. Despite an extensive description of the spore wall, little is known regarding the mechanism by which it is deposited or the role it plays in cell adhesion and infection. In this study, we report the identification and characterization of two novel spore wall proteins, SWP7 and SWP9, in the microsporidian speciesNosema bombycis. SWP7 and SWP9 are mainly localized to the exospore and endospore of mature spores and the cytoplasm of sporonts, respectively. In addition, a portion of SWP9 is targeted to the spore wall of sporoblasts earlier than SWP7 is. Both SWP7 and SWP9 are specifically colocalized to the spore wall in mature spores. Furthermore, immunoprecipitation, far-Western blotting, unreduced SDS-PAGE, and yeast two-hybrid data demonstrated that SWP7 interacted with SWP9. The chitin binding assay showed that, within the total spore protein, SWP9 and SWP7 can bind to the deproteinated chitin spore coats (DCSCs) ofN. bombycis. However, binding of the recombinant protein rSWP7-His to the DCSCs is dependent on the combination of rSWP9–glutathioneS-transferase (GST) with the DCSCs. Finally, rSWP9-GST, anti-SWP9, and anti-SWP7 antibodies decreased spore adhesion and infection of the host cell. In conclusion, SWP7 and SWP9 may have important structural capacities and play significant roles in modulating host cell adherence and infectionin vitro. A possible major function of SWP9 is as a scaffolding protein that supports other proteins (such as SWP7) that form the integrated spore wall ofN. bombycis.

scholarly journals SWP5, a Spore Wall Protein, Interacts with Polar Tube Proteins in the Parasitic Microsporidian Nosema bombycis

2011 ◽  
Vol 11 (2) ◽  
pp. 229-237 ◽  
Author(s):  
Zhi Li ◽  
Guoqing Pan ◽  
Tian Li ◽  
Wei Huang ◽  
Jie Chen ◽  
...  
Keyword(s):  
Structural Integrity ◽  
Sequence Similarity ◽  
Spore Wall ◽  
Host Cells ◽  
Polar Tube ◽  
Content Type ◽  
Nosema Bombycis ◽  
Encephalitozoon Intestinalis ◽  
Intracellular Parasites ◽  
Almost All

ABSTRACTMicrosporidia are a group of eukaryotic intracellular parasites that infect almost all vertebrates and invertebrates. The microsporidian invasion process involves the extrusion of a unique polar tube into host cells. Both the spore wall and the polar tube play an important role in microsporidian pathogenesis. So far, five spore wall proteins (SWP1, SWP2, Enp1, Enp2, and EcCDA) fromEncephalitozoon intestinalisandEncephalitozoon cuniculiand five spore wall proteins (SWP32, SWP30, SWP26, SWP25, and NbSWP5) from the silkworm pathogenNosema bombycishave been identified. Here we report the identification and characterization of a spore wall protein (SWP5) with a molecular mass of 20.3 kDa inN. bombycis. This protein has low sequence similarity to other eukaryotic proteins. Immunolocalization analysis showed SWP5 localized to the exospore and the region of the polar tube in mature spores. Immunoprecipitation, mass spectrometry, and immunofluorescence analyses revealed that SWP5 interacts with the polar tube proteins PTP2 and PTP3. Anti-SWP5 serum pretreatment of mature spores significantly decreased their polar tube extrusion rate. Taken together, our results show that SWP5 is a spore wall protein localized to the spore wall and that it interacts with the polar tube, may play an important role in supporting the structural integrity of the spore wall, and potentially modulates the course of infection ofN. bombycis.

scholarly journals Chitinase 3-Like 1 Promotes Candida albicans Killing and Preserves Corneal Structure and Function by Controlling Host Antifungal Responses

2015 ◽  
Vol 83 (10) ◽  
pp. 4154-4164 ◽  
Author(s):  
Nan Gao ◽  
Fu-Shin X. Yu
Keyword(s):  
Candida Albicans ◽  
Tissue Injury ◽  
Fungal Keratitis ◽  
Innate Immune ◽  
Therapeutic Window ◽  
Minor Role ◽  
Dependent Manner ◽  
Content Type ◽  
And Function ◽  
Interfering Rna

Chitinase 3-like 1 (CHI3L1) has been shown to play a role in promoting antibacterial responses, decreasing tissue injury, and enhancing pulmonary repair. This study sought to elucidate the role of CHI3L1 in augmenting the corneal innate immune response toCandida albicansinfection in an animal model of fungal keratitis. Flagellin applied topically 24 h prior toC. albicansinoculation significantly protected the corneal fromC. albicansand induced CHI3L1 expression in C57BL/6 mouse corneas. CHI3L1, however, played a detectable but minor role in flagellin-induced protection. WhileC. albicanskeratitis was more severe in the corneas treated withChi3l1small interfering RNA (siRNA), corneas treated with recombinant CHI3L1 beforeC. albicansinoculation had markedly ameliorated keratitis, reduced fungal load, and decreased polymorphonucleocyte (PMN) infiltration in an interleukin 13 receptor α2 (IL-13Rα2)-dependent manner. CHI3L1 treatment resulted in the induction of the antimicrobial peptides β-defensin 3, CRAMP, and chemokine CXCL10 and its receptor CXCR3 in corneal epithelial cells. Importantly, CHI3L1 administered afterC. albicansinoculation also had strong protection against fungal keratitis, suggesting a therapeutic window. This is the first report demonstrating that CHI3L1 is induced during fungal infection, where it acts as an immunomodulator to promote fungal clearance and to regulate antifungal innate immune responses in the cornea.

scholarly journals Interactions of Encephalitozoon cuniculi Polar Tube Proteins

2010 ◽  
Vol 78 (6) ◽  
pp. 2745-2753 ◽  
Author(s):  
Boumediene Bouzahzah ◽  
Fnu Nagajyothi ◽  
Kaya Ghosh ◽  
Peter M. Takvorian ◽  
Ann Cali ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Host Cells ◽  
Encephalitozoon Cuniculi ◽  
Polar Tube ◽  
Yeast Two Hybrid ◽  
Obligate Intracellular ◽  
Repetitive Motif ◽  
And Function ◽  
Two Hybrid

ABSTRACT Microsporidia are eukaryotic, obligate intracellular organisms defined by small spores that contain a single invasion organelle, the polar tube, which coils around the interior of the spore. When these parasites infect host cells, the polar tube is discharged from the anterior pole of the spore, pierces the cell, and transfers sporoplasm into the cytoplasm of the host. Three polar tube proteins (PTP1, PTP2, and PTP3) have been identified in this structure. The interactions of these proteins in the assembly and function of the polar tube are not known. This study was undertaken to examine the protein interactions of the Encephalitozoon cuniculi polar tube proteins (EcPTPs). Immunofluorescence and immunoelectron microscopy confirmed the colocalization of EcPTP1, EcPTP2, and EcPTP3 to the polar tube. Experiments using cross-linkers indicated that EcPTP1, EcPTP2, and EcPTP3 form a complex in the polar tube, which was confirmed by immunoprecipitation using EcPTP1 antiserum. Yeast two-hybrid analysis revealed that full-length EcPTP1, EcPTP2, and EcPTP3 interact with each other in vivo. Both the N and C termini of EcPTP1 were involved in these interactions, but the central region of this protein, which contains a repetitive motif, was not. Further studies of polar tube proteins and their structural interactions may help elucidate the formation of the polar tube during the invasion process.

scholarly journals Proteomics Screen Identifies Class I Rab11 Family Interacting Proteins as Key Regulators of Cytokinesis

2016 ◽  
Vol 37 (3) ◽  
Author(s):  
Carl Laflamme ◽  
Jacob A. Galan ◽  
Khaled Ben El Kadhi ◽  
Antoine Méant ◽  
Carlos Zeledon ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Protein Trafficking ◽  
Protein Transport ◽  
Effector Proteins ◽  
Class I ◽  
Dependent Manner ◽  
Interacting Proteins ◽  
Content Type ◽  
Evolutionarily Conserved ◽  
Dependent Protein

ABSTRACTThe 14-3-3 protein family orchestrates a complex network of molecular interactions that regulates various biological processes. Owing to their role in regulating the cell cycle and protein trafficking, 14-3-3 proteins are prevalent in human diseases such as cancer, diabetes, and neurodegeneration. 14-3-3 proteins are expressed in all eukaryotic cells, suggesting that they mediate their biological functions through evolutionarily conserved protein interactions. To identify these core 14-3-3 client proteins, we used an affinity-based proteomics approach to characterize and compare the human andDrosophila14-3-3 interactomes. Using this approach, we identified a group of Rab11 effector proteins, termed class I Rab11 family interacting proteins (Rab11-FIPs), or Rip11 inDrosophila. We found that 14-3-3 binds to Rip11 in a phospho-dependent manner to ensure its proper subcellular distribution during cell division. Our results indicate that Rip11 plays an essential role in the regulation of cytokinesis and that this function requires its association with 14-3-3 but not with Rab11. Together, our results suggest an evolutionarily conserved role for 14-3-3 in controlling Rip11-dependent protein transport during cytokinesis.

EFEITOS DO DEEP RUNNING SOBRE A AMPLITUDE DE MOVIMENTO E A CAPACIDADE FUNCIONAL DOS MEMBROS SUPERIORES DE MULHERES MASTECTOMIZADAS: RELATO DE CASOS

2019 ◽  
pp. 121-131
Keyword(s):  
Range Of Motion ◽  
Upper Limb ◽  
Functional Capacity ◽  
External Rotation ◽  
Treatment Protocol ◽  
Range Of Movement ◽  
Flexion Extension ◽  
Before And After ◽  
In The Beginning ◽  
And Function

Introduction: Breast cancer is the most common type of cancer among women in Brazil and in the worl. The surgical treatment procedure may cause severe morbidity in the upper limb homolateral to surgery, including the reduction of the range of motion, with consequent impairment of function. A physiotherapeutic approach has an important role in the recover range of motion and the functionality of these women, guaranteeing the occupational, domestestic, familiar and conjugated activities, and, in this way, also improving the quality of life. Objectives: To analyse chances in the shoulder's range of motion and the functional capacity of the upper limbs, promoted by the deep running procedure in women with late postoperative mastectomy. Methods: All the patients were submitted to an evaluation in the beginning and end of the treatment, including: goniometry of flexion, extension, abduction, adduction, internal and external rotation of the shoulder joint; and function capacity analysis in activities that involve the upper members by DASH questionnaire. The treatment protocol includes twelve sessions of deep running, realized twice a week, in deep pool, for 20-minute during six weeks. Results: Were submitted to treatment a total of 4 patients. Despite the improvement in the numerical values, statistically significant differences were not found on the range of movements and in the functional capacity of upper members before and after the deep running sessions in post-mastectomy women. Conclusion: Deep running had effects on the numerical values of range of movement and upper limb functionality in women in the late postoperative period of the mastectomy procedure, but without statistically significant differences.

scholarly journals In VitroAnalyses of the Effects of Heparin and Parabens on Candida albicans Biofilms and Planktonic Cells

2011 ◽  
Vol 56 (1) ◽  
pp. 148-153 ◽  
Author(s):  
Marisa H. Miceli ◽  
Stella M. Bernardo ◽  
T. S. Neil Ku ◽  
Carla Walraven ◽  
Samuel A. Lee
Keyword(s):  
Candida Albicans ◽  
Metabolic Activity ◽  
Biofilm Formation ◽  
High Dose ◽  
Dependent Manner ◽  
Planktonic Cells ◽  
Content Type ◽  
Heparin Sodium ◽  
The Individual

ABSTRACTInfections and thromboses are the most common complications associated with central venous catheters. Suggested strategies for prevention and management of these complications include the use of heparin-coated catheters, heparin locks, and antimicrobial lock therapy. However, the effects of heparin onCandida albicansbiofilms and planktonic cells have not been previously studied. Therefore, we sought to determine thein vitroeffect of a heparin sodium preparation (HP) on biofilms and planktonic cells ofC. albicans. Because HP contains two preservatives, methyl paraben (MP) and propyl paraben (PP), these compounds and heparin sodium without preservatives (Pure-H) were also tested individually. The metabolic activity of the mature biofilm after treatment was assessed using XTT [2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] reduction and microscopy. Pure-H, MP, and PP caused up to 75, 85, and 60% reductions of metabolic activity of the mature preformedC. albicansbiofilms, respectively. Maximal efficacy against the mature biofilm was observed with HP (up to 90%) compared to the individual compounds (P< 0.0001). Pure-H, MP, and PP each inhibitedC. albicansbiofilm formation up to 90%. A complete inhibition of biofilm formation was observed with HP at 5,000 U/ml and higher. When tested against planktonic cells, each compound inhibited growth in a dose-dependent manner. These data indicated that HP, MP, PP, and Pure-H havein vitroantifungal activity againstC. albicansmature biofilms, formation of biofilms, and planktonic cells. Investigation of high-dose heparin-based strategies (e.g., heparin locks) in combination with traditional antifungal agents for the treatment and/or prevention ofC. albicansbiofilms is warranted.

scholarly journals Identification and Biochemical Characterization of High Mobility Group Protein 20A as a Novel Ca2+/S100A6 Target

Biomolecules ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 510
Author(s):  
Maho Yamamoto ◽  
Rina Kondo ◽  
Haruka Hozumi ◽  
Seita Doi ◽  
Miwako Denda ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Biochemical Characterization ◽  
S100 Protein ◽  
High Mobility ◽  
Dependent Manner ◽  
Protein Signaling ◽  
Protein Protein Interactions ◽  
High Mobility Group Protein ◽  
Pulldown Assay

During screening of protein-protein interactions, using human protein arrays carrying 19,676 recombinant glutathione s-transferase (GST)-fused human proteins, we identified the high-mobility protein group 20A (HMG20A) as a novel S100A6 binding partner. We confirmed the Ca2+-dependent interaction of HMG20A with S100A6 by the protein array method, biotinylated S100A6 overlay, and GST-pulldown assay in vitro and in transfected COS-7 cells. Co-immunoprecipitation of S100A6 with HMG20A from HeLa cells in a Ca2+-dependent manner revealed the physiological relevance of the S100A6/HMG20A interaction. In addition, HMG20A has the ability to interact with S100A1, S100A2, and S100B in a Ca2+-dependent manner, but not with S100A4, A11, A12, and calmodulin. S100A6 binding experiments using various HMG20A mutants revealed that Ca2+/S100A6 interacts with the C-terminal region (residues 311–342) of HMG20A with stoichiometric binding (HMG20A:S100A6 dimer = 1:1). This was confirmed by the fact that a GST-HMG20A mutant lacking the S100A6 binding region (residues 311–347, HMG20A-ΔC) failed to interact with endogenous S100A6 in transfected COS-7 cells, unlike wild-type HMG20A. Taken together, these results identify, for the first time, HMG20A as a target of Ca2+/S100 proteins, and may suggest a novel linkage between Ca2+/S100 protein signaling and HMG20A function, including in the regulation of neural differentiation.

scholarly journals C. elegans germ granules require both assembly and localized regulators for mRNA repression

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Scott Takeo Aoki ◽  
Tina R. Lynch ◽  
Sarah L. Crittenden ◽  
Craig A. Bingman ◽  
Marvin Wickens ◽  
...  
Keyword(s):  
Liquid Droplet ◽  
Scaffolding Protein ◽  
P Granules ◽  
C Elegans ◽  
Cytoplasmic Rna ◽  
And Function ◽  
Rnp Granules ◽  
Key Residues ◽  
Reporter Mrna

AbstractCytoplasmic RNA–protein (RNP) granules have diverse biophysical properties, from liquid to solid, and play enigmatic roles in RNA metabolism. Nematode P granules are paradigmatic liquid droplet granules and central to germ cell development. Here we analyze a key P granule scaffolding protein, PGL-1, to investigate the functional relationship between P granule assembly and function. Using a protein–RNA tethering assay, we find that reporter mRNA expression is repressed when recruited to PGL-1. We determine the crystal structure of the PGL-1 N-terminal region to 1.5 Å, discover its dimerization, and identify key residues at the dimer interface. Mutations of those interface residues prevent P granule assembly in vivo, de-repress PGL-1 tethered mRNA, and reduce fertility. Therefore, PGL-1 dimerization lies at the heart of both P granule assembly and function. Finally, we identify the P granule-associated Argonaute WAGO-1 as crucial for repression of PGL-1 tethered mRNA. We conclude that P granule function requires both assembly and localized regulators.

scholarly journals The role of NbTMP1, a surface protein of sporoplasm, in Nosema bombycis infection

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Shiyi Zheng ◽  
Yukang Huang ◽  
Hongyun Huang ◽  
Bin Yu ◽  
Ni Zhou ◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Selective Breeding ◽  
Developmental Stages ◽  
Transmembrane Protein ◽  
Surface Protein ◽  
Spore Wall ◽  
Significant Inhibition ◽  
Pcr Analysis ◽  
Immunofluorescence Analysis ◽  
Nosema Bombycis

Abstract Background Nosema bombycis is a unicellular eukaryotic pathogen of the silkworm, Bombyx mori, and is an economic and occupational hazard in the silkworm industry. Because of its long incubation period and horizontal and vertical transmission, it is subject to quarantine measures in sericulture production. The microsporidian life-cycle includes a dormant extracellular phase and intracellular proliferation phase, with the proliferation period being the most active period. This latter period lacks spore wall protection and may be the most susceptible stage for control. Methods In order to find suitable target for the selective breeding of N. bombycis-resistant silkworm strains, we screen highly expressed membrane proteins from the transcriptome data of N. bombycis. The subcellular localization of the candidate protein was verified by Indirect immunofluorescence analysis (IFA) and immunoelectron microscopy (IEM), and its role in N. bombycis proliferation was verified by RNAi. Results The N. bombycis protein (NBO_76g0014) was identified as a transmembrane protein and named NbTMP1. It is homologous with hypothetical proteins NGRA_1734 from Nosema granulosis. NbTMP1 has a transmembrane region of 23 amino acids at the N-terminus. Indirect immunofluorescence analysis (IFA) results suggest that NbTMP1 is secreted on the plasma membrane as the spores develop. Western blot and qRT-PCR analysis showed that NbTMP1 was expressed in all developmental stages of N. bombycis in infected cells and in the silkworm midgut. Downregulation of NbTMP1 expression resulted in significant inhibition of N. bombycis proliferation. Conclusions We confirmed that NbTMP1 is a membrane protein of N. bombycis. Reduction of the transcription level of NbTMP1 significantly inhibited N. bombycis proliferation, and this protein may be a target for the selective breeding of N. bombycis-resistant silkworm strains.

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