scholarly journals First molecular insights into the infection process provoked by Neofusicoccum parvum in Liquidambar styraciflua and the identification of new cysteine-rich proteins in both organisms

2020 ◽  
Author(s):  
Rebeca Vázquez-Avendaño ◽  
José Benjamín Rodríguez-Haas ◽  
Hugo Velázquez-Delgado ◽  
Greta Hanako Rosas-Saito ◽  
Eric Edmundo Hernández-Domínguez ◽  
...  
Keyword(s):  
Post Infection ◽  
Molecular Mechanisms ◽  
Phylogenetic Analyses ◽  
Leaf Tissue ◽  
Infection Process ◽  
Liquidambar Styraciflua ◽  
Broad Host Range ◽  
Neofusicoccum Parvum ◽  
First Time

Abstract Background Neofusicoccum parvum belongs to the Botryosphaeriaceae family, which groups endophytic and latent pathogens of woody plants responsible for diseases such as cankers, dieback and blight. It is a widespread pathogen with a broad host range, including agricultural, horticultural and forestry plants; therefore, it is relevant to characterize the molecular mechanisms involved in the disease caused by this pathogen. This work reports for the first time N. parvum as a pathogen of Liquidambar styraciflua. We established an in vitro pathosystem using foliar tissue to characterize the infection process through scanning electron microscopy (SEM). Because cysteine-rich proteins (CysRPs) have been studied for their important functions in plant-pathogen interactions, new CysRPs were identified for these organisms, and mRNA expression of these proteins was analyzed at early time points during the interaction.Results 24 hours post infection, the pathogen caused visible symptoms, and microscopic analysis at 16 days post infection revealed the presence of N. parvum pycnidia embedded in L. styraciflua leaf tissue. For both organisms, two databases with transcriptomic and genomic information were analyzed, and five new CysRPs were identified for each organism. The length varied between 95 and 204 amino acids, and in silico analysis revealed that all the proteins are potentially secreted. The search for conserved domains and phylogenetic analyses revealed that all the proteins are novel, including two of N. parvum that present the well-known CFEM domain. RT-qPCR analysis was conducted at 24 and 72 hours post infection, and the results showed changes in the levels of CysRP mRNAs for both the plant and the fungus at early stages during the interaction.Conclusions N. parvum was identified for the first time as a pathogen of L. styraciflua, and this work presents an approach to comprehensively understand the molecular mechanisms involved in this interaction, highlighting the potential involvement of CysRPs of both organisms under this biotic stress.

scholarly journals First molecular insights into the infection process provoked by Neofusicoccum parvum in Liquidambar styraciflua and the identification of new cysteine rich proteins in both organisms

2020 ◽  
Author(s):  
Rebeca Vázquez-Avendaño ◽  
José Benjamín Rodríguez-Haas ◽  
Hugo Velázquez-Delgado ◽  
Greta Hanako Rosas-Saito ◽  
Eric Edmundo Hernández-Domínguez ◽  
...  
Keyword(s):  
Post Infection ◽  
Molecular Mechanisms ◽  
Phylogenetic Analyses ◽  
Leaf Tissue ◽  
Infection Process ◽  
Liquidambar Styraciflua ◽  
Broad Host Range ◽  
Neofusicoccum Parvum ◽  
First Time

Abstract Background Neofusicoccum parvum belongs to Botryosphaeriaceae family that groups endophytic and latent pathogens of woody plants responsible of diseases such as cankers, dieback and blight. Is a widespread pathogen in a broad host range including agricultural, horticultural and forestry plants, therefore is relevant to characterize molecular mechanisms involved in the disease. This work, report for first time a N. parvum as a pathogen of Liquidambar styraciflua. We established an in vitro pathosystem using foliar tissue in order to characterize the infection process through scanning electron microscopy. Because cysteine rich proteins (CysRPs) are well described for their important functions under plant-pathogen interaction, new CysRPs were identified for these organisms, and mRNAs expression of these proteins was analyzed at early times during the interaction.Results Since the first 24 hours post infection, the pathogen caused visible symptoms and the microscopic analysis at 16 days post infection revealed the presence of N. parvum pycnidium immersed in L. styraciflua leaf tissue. For both organisms, two databases with transcriptomic and genomic information were analyzed and new five CysRPs were identified for each organism, the amino acid length varied between 95 and 204 and in silico analysis revealed that all proteins are potentially secreted. The search of conserved domains and phylogenetic analyses bring to light that all proteins are novel including two of N. parvum that present the well-known CFEM domain. RT-qPCR analysis was conducted at 24- and 72-hours post infection and the results showed change levels of CysRPs mRNAs for both the plant and the fungus at early times during the interaction.ConclusionsIt is recognized for first time that N. parvum is a pathogen of L. styraciflua and this work represents an approach to begin to deeply understand the molecular mechanisms involved in this interaction highlighting the potential involvement of CysRPs of both organism during this biotic stress.

scholarly journals First molecular insights into the infection process provoked by Neofusicoccum parvum in Liquidambar styraciflua and the identification of new cysteine-rich proteins in both organisms

2020 ◽  
Author(s):  
Rebeca Vázquez-Avendaño ◽  
José Benjamín Rodríguez-Haas ◽  
Hugo Velázquez-Delgado ◽  
Greta Hanako Rosas-Saito ◽  
Eric Edmundo Hernández-Domínguez ◽  
...  
Keyword(s):  
Post Infection ◽  
Molecular Mechanisms ◽  
Phylogenetic Analyses ◽  
Leaf Tissue ◽  
Infection Process ◽  
Liquidambar Styraciflua ◽  
Broad Host Range ◽  
Neofusicoccum Parvum ◽  
First Time

Abstract Background: Neofusicoccum parvum belongs to the Botryosphaeriaceae family, which groups endophytic and latent pathogens of woody plants responsible for diseases such as cankers, dieback and blight. It is a widespread pathogen with a broad host range, including agricultural, horticultural and forestry plants; therefore, it is relevant to characterize the molecular mechanisms involved in the disease caused by this pathogen. This work reports for the first time N. parvum as a pathogen of Liquidambar styraciflua. We established an in vitro pathosystem using foliar tissue to characterize the infection process through scanning electron microscopy (SEM). Because cysteine-rich proteins (CysRPs) have been studied for their important functions in plant-pathogen interactions, new CysRPs were identified for these organisms, and mRNA expression of these proteins was analyzed at early time points during the interaction.Results: After the first 24 hours post infection, the pathogen caused visible symptoms, and microscopic analysis at 16 days post infection revealed the presence of N. parvum pycnidia embedded in L. styraciflua leaf tissue. For both organisms, two databases with transcriptomic and genomic information were analyzed, and five new CysRPs were identified for each organism. The length varied between 95 and 204 amino acids, and in silico analysis revealed that all the proteins are potentially secreted. The search for conserved domains and phylogenetic analyses revealed that all the proteins are novel, including two of N. parvum that present the well-known CFEM domain. RT-qPCR analysis was conducted at 24 and 72 hours post infection, and the results showed changes in the levels of CysRP mRNAs for both the plant and the fungus at early stages during the interaction.Conclusions: N. parvum was identified for the first time as a pathogen of L. styraciflua, and this work presents an approach to comprehensively understand the molecular mechanisms involved in this interaction, highlighting the potential involvement of CysRPs of both organisms under this biotic stress.

scholarly journals Insights of the Neofusicoccum parvum–Liquidambar styraciflua Interaction and Identification of New Cysteine-Rich Proteins in Both Species

2021 ◽  
Vol 7 (12) ◽  
pp. 1027
Author(s):  
Rebeca Vázquez-Avendaño ◽  
José Benjamín Rodríguez-Haas ◽  
Hugo Velázquez-Delgado ◽  
Greta Hanako Rosas-Saito ◽  
Eric Edmundo Hernández-Domínguez ◽  
...  
Keyword(s):  
In Silico Analysis ◽  
Infection Process ◽  
Liquidambar Styraciflua ◽  
Genomic Databases ◽  
Neofusicoccum Parvum ◽  
Disease Symptoms ◽  
Foliar Tissue ◽  
First Time ◽  
Silico Analysis

Neofusicoccum parvum belongs to the Botryosphaeriaceae family, which contains endophytes and pathogens of woody plants. In this study, we isolated 11 strains from diseased tissue of Liquidambar styraciflua. Testing with Koch’s postulates—followed by a molecular approach—revealed that N. parvum was the most pathogenic strain. We established an in vitro pathosystem (L. styraciflua foliar tissue–N. parvum) in order to characterize the infection process during the first 16 days. New CysRPs were identified for both organisms using public transcriptomic and genomic databases, while mRNA expression of CysRPs was analyzed by RT-qPCR. The results showed that N. parvum caused disease symptoms after 24 h that intensified over time. Through in silico analysis, 5 CysRPs were identified for each organism, revealing that all of the proteins are potentially secreted and novel, including two of N. parvum proteins containing the CFEM domain. Interestingly, the levels of the CysRPs mRNAs change during the interaction. This study reports N. parvum as a pathogen of L. styraciflua for the first time and highlights the potential involvement of CysRPs in both organisms during this interaction.

scholarly journals Analysis of a Preliminary microRNA Expression Signature in a Human Telangiectatic Osteogenic Sarcoma Cancer Cell Line

2021 ◽  
Vol 22 (3) ◽  
pp. 1163
Author(s):  
Gaia Palmini ◽  
Cecilia Romagnoli ◽  
Simone Donati ◽  
Roberto Zonefrati ◽  
Gianna Galli ◽  
...  
Keyword(s):  
Cell Line ◽  
Molecular Mechanisms ◽  
Osteogenic Sarcoma ◽  
Cancer Cell Line ◽  
Microscopic Features ◽  
In Vitro Establishment ◽  
Profile Characteristic ◽  
First Time

Telangiectatic osteosarcoma (TOS) is an aggressive variant of osteosarcoma (OS) with distinctive radiographic, gross, microscopic features, and prognostic implications. Despite several studies on OS, we are still far from understanding the molecular mechanisms of TOS. In recent years, many studies have demonstrated not only that microRNAs (miRNAs) are involved in OS tumorigenesis, development, and metastasis, but also that the presence in high-grade types of OS of cancer stem cells (CSCs) plays an important role in tumor progression. Despite these findings, nothing has been described previously about the expression of miRNAs and the presence of CSCs in human TOS. Therefore, we have isolated/characterized a putative CSC cell line from human TOS (TOS-CSCs) and evaluated the expression levels of several miRNAs in TOS-CSCs using real-time quantitative assays. We show, for the first time, the existence of CSCs in human TOS, highlighting the in vitro establishment of this unique stabilized cell line and an identification of a preliminary expression of the miRNA profile, characteristic of TOS-CSCs. These findings represent an important step in the study of the biology of one of the most aggressive variants of OS and the role of miRNAs in TOS-CSC behavior.

scholarly journals A Central Role for the Armadillo Protein Plakoglobin in the Autoimmune Disease Pemphigus Vulgaris

2001 ◽  
Vol 153 (4) ◽  
pp. 823-834 ◽  
Author(s):  
Reto Caldelari ◽  
Alain de Bruin ◽  
Dominique Baumann ◽  
Maja M. Suter ◽  
Christiane Bierkamp ◽  
...  
Keyword(s):  
Steric Hindrance ◽  
In Vitro Model ◽  
Molecular Mechanisms ◽  
Pemphigus Vulgaris ◽  
Cell Types ◽  
Linear Distribution ◽  
Igg Binding ◽  
Vitro Model ◽  
First Time

In pemphigus vulgaris (PV), autoantibody binding to desmoglein (Dsg) 3 induces loss of intercellular adhesion in skin and mucous membranes. Two hypotheses are currently favored to explain the underlying molecular mechanisms: (a) disruption of adhesion through steric hindrance, and (b) interference of desmosomal cadherin-bound antibody with intracellular events, which we speculated to involve plakoglobin. To investigate the second hypothesis we established keratinocyte cultures from plakoglobin knockout (PG−/−) embryos and PG+/+ control mice. Although both cell types exhibited desmosomal cadherin-mediated adhesion during calcium-induced differentiation and bound PV immunoglobin (IgG) at their cell surface, only PG+/+ keratinocytes responded with keratin retraction and loss of adhesion. When full-length plakoglobin was reintroduced into PG−/− cells, responsiveness to PV IgG was restored. Moreover, in these cells like in PG+/+ keratinocytes, PV IgG binding severely affected the linear distribution of plakoglobin at the plasma membrane. Taken together, the establishment of an in vitro model using PG+/+ and PG−/− keratinocytes allowed us (a) to exclude the steric hindrance only hypothesis, and (b) to demonstrate for the first time that plakoglobin plays a central role in PV, a finding that will provide a novel direction for investigations of the molecular mechanisms leading to PV, and on the function of plakoglobin in differentiating keratinocytes.

scholarly journals Transcriptomic Analysis of mRNA-lncRNA-miRNA Interactions in Hepatocellular Carcinoma

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Xia Tang ◽  
Delong Feng ◽  
Min Li ◽  
Jinxue Zhou ◽  
Xiaoyuan Li ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Molecular Mechanisms ◽  
Expression Profiles ◽  
Rna Seq ◽  
Pairwise Interactions ◽  
Micro Rnas ◽  
Non Coding Rnas ◽  
First Time

Abstract Fully elucidating the molecular mechanisms of non-coding RNAs (ncRNAs), including micro RNAs (miRNAs) and long non-coding RNAs (lncRNAs), underlying hepatocarcinogenesis is challenging. We characterized the expression profiles of ncRNAs and constructed a regulatory mRNA-lncRNA-miRNA (MLMI) network based on transcriptome sequencing (RNA-seq) of hepatocellular carcinoma (HCC, n = 9) patients. Of the identified miRNAs (n = 203) and lncRNAs (n = 1,090), we found 16 significantly differentially expressed (DE) miRNAs and three DE lncRNAs. The DE RNAs were highly enriched in 21 functional pathways implicated in HCC (p < 0.05), including p53, MAPK, and NAFLD signaling. Potential pairwise interactions between DE ncRNAs and mRNAs were fully characterized using in silico prediction and experimentally-validated evidence. We for the first time constructed a MLMI network of reciprocal interactions for 16 miRNAs, three lncRNAs, and 253 mRNAs in HCC. The predominant role of MEG3 in the MLMI network was validated by its overexpression in vitro that the expression levels of a proportion of MEG3-targeted miRNAs and mRNAs was changed significantly. Our results suggested that the comprehensive MLMI network synergistically modulated carcinogenesis, and the crosstalk of the network provides a new avenue to accurately describe the molecular mechanisms of hepatocarcinogenesis.

scholarly journals Chemerin-ChemR23 Signaling in Tooth Development

2012 ◽  
Vol 91 (12) ◽  
pp. 1147-1153 ◽  
Author(s):  
T. Ohira ◽  
D. Spear ◽  
N. Azimi ◽  
V. Andreeva ◽  
P.C. Yelick
Keyword(s):  
Progenitor Cells ◽  
Molecular Mechanisms ◽  
Tooth Development ◽  
Expression Patterns ◽  
Cell Interactions ◽  
Specific Expression ◽  
Ribosomal Protein S6 ◽  
First Time

Our long-term goal is to identify and characterize molecular mechanisms regulating tooth development, including those mediating the critical dental epithelial-dental mesenchymal (DE-DM) cell interactions required for normal tooth development. The goal of this study was to investigate Chemerin (Rarres2)/ChemR23(Cmklr1) signaling in DE-DM cell interactions in normal tooth development. Here we present, for the first time, tissue-specific expression patterns of Chemerin and ChemR23 in mouse tooth development. We show that Chemerin is expressed in cultured DE progenitor cells, while ChemR23 is expressed in cultured DM cells. Moreover, we demonstrate that ribosomal protein S6 (rS6) and Akt, downstream targets of Chemerin/ChemR23 signaling, are phosphorylated in response to Chemerin/ChemR23 signaling in vitro and are expressed in mouse tooth development. Together, these results suggest roles for Chemerin/ChemR23-mediated DE-DM cell signaling during tooth morphogenesis.

scholarly journals P4HA2 Promotes Epithelial-to-Mesenchymal Transition and Glioma Malignancy through the Collagen-Dependent PI3K/AKT Pathway

2021 ◽  
Vol 2021 ◽  
pp. 1-14
Author(s):  
Jing Lin ◽  
Lei Jiang ◽  
Xiaogang Wang ◽  
Wenxin Wei ◽  
Chaoli Song ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Epithelial To Mesenchymal Transition ◽  
Enzyme Inhibitor ◽  
Tumor Xenograft ◽  
Mesenchymal Transition ◽  
Akt Activation ◽  
Collagen Genes ◽  
First Time

Prolyl-4-hydroxylase subunit 2 (P4HA2) is a member of collagen modification enzymes involved in the remodeling of the extracellular matrix (ECM). Mounting evidence has suggested that deregulation of P4HA2 is common in cancer. However, the role of P4HA2 in glioma remains unknown. The present study aimed to elucidate the expression pattern, oncogenic functions, and molecular mechanisms of P4HA2 in glioblastoma cells. The TCGA datasets and paraffin samples were used for examining the expressions of P4HA2. P4HA2-specific lentivirus was generated to assess its oncogenic functions. A P4HA2 enzyme inhibitor (DHB) and an AKT agonist (SC79) were utilized to study the mechanisms. As a result, we demonstrated that P4HA2 is overexpressed in glioma and inversely correlates with patient survival. Knockdown of P4HA2 inhibited proliferation, migration, invasion, and epithelial-to-mesenchymal transition (EMT) like phenotype of glioma cells in vitro and suppressed tumor xenograft growth in vivo. Mechanistically, expressions of a series of collagen genes and of phosphorylated PI3K/AKT were downregulated by either P4HA2 silencing or inhibition of its prolyl hydroxylase. Finally, the inhibitory effects on the migration, invasion, and EMT-related molecules by P4HA2 knockdown were reversed by AKT activation with SC79. Our findings for the first time reveal that P4HA2 acts as an oncogenic molecule in glioma malignancy by regulating the expressions of collagens and the downstream PI3K/AKT signaling pathway.

scholarly journals A SilencedvanAGene Cluster on a Transferable Plasmid Caused an Outbreak of Vancomycin-Variable Enterococci

2016 ◽  
Vol 60 (7) ◽  
pp. 4119-4127 ◽  
Author(s):  
Audun Sivertsen ◽  
Torunn Pedersen ◽  
Kjersti Wik Larssen ◽  
Kåre Bergh ◽  
Torunn Gresdal Rønning ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Molecular Mechanisms ◽  
Genetic Difference ◽  
Brain Heart Infusion ◽  
Broad Host Range ◽  
S1 Nuclease ◽  
Content Type ◽  
Transcription Profiles

ABSTRACTWe report an outbreak of vancomycin-variablevanA+enterococci (VVE) able to escape phenotypic detection by current guidelines and demonstrate the molecular mechanisms forin vivoswitching into vancomycin resistance and horizontal spread of thevanAcluster. Forty-eightvanA+Enterococcus faeciumisolates and oneEnterococcus faecalisisolate were analyzed for clonality with pulsed-field gel electrophoresis (PFGE), and theirvanAgene cluster compositions were assessed by PCR and whole-genome sequencing of six isolates. The susceptible VVE strains were cultivated in brain heart infusion broth containing vancomycin at 8 μg/ml forin vitrodevelopment of resistant VVE. The transcription profiles of susceptible VVE and their resistant revertants were assessed using quantitative reverse transcription-PCR. Plasmid content was analyzed with S1 nuclease PFGE and hybridizations. Conjugative transfer ofvanAwas assessed by filter mating. The only genetic difference between thevanAclusters of susceptible and resistant VVE was an ISL3-family element upstream ofvanHAX, which silencedvanHAXgene transcription in susceptible VVE. Furthermore, the VVE had an insertion of IS1542betweenorf2andvanRthat attenuated the expression ofvanHAX. Growth of susceptible VVE occurred after 24 to 72 h of exposure to vancomycin due to excision of the ISL3-family element. ThevanAgene cluster was located on a transferable broad-host-range plasmid also detected in outbreak isolates with different pulsotypes, including oneE. faecalisisolate. Horizontally transferable silencedvanAable to escape detection and revert into resistance during vancomycin therapy represents a new challenge in the clinic. Genotypic testing of invasive vancomycin-susceptible enterococci byvanA-PCR is advised.

scholarly journals LtEPG1, a Secretory Endopolygalacturonase Protein, Regulates the Virulence of Lasiodiplodia theobromae in Vitis vinifera and Is Recognized as a Microbe-Associated Molecular Patterns

2020 ◽  
Vol 110 (10) ◽  
pp. 1727-1736
Author(s):  
K. W. Thilini Chethana ◽  
Junbo Peng ◽  
Xinghong Li ◽  
Qikai Xing ◽  
Mei Liu ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Transcript Level ◽  
Infection Process ◽  
Glycoside Hydrolases ◽  
Glycoside Hydrolase Family ◽  
In Planta ◽  
Lasiodiplodia Theobromae ◽  
Successful Infection ◽  
Molecular Patterns

The Lasiodiplodia theobromae genome encodes numerous glycoside hydrolases involved in organic matter degradation and conducive to pathogen infection, whereas their molecular mechanisms are still largely unknown. Here, we identified the glycoside hydrolase family 28 endopolygalacturonase LtEPG1 in L. theobromae and characterized its function in detail. LtEPG1 acts as a virulence factor during L. theobromae infection. Overexpression and silencing of LtEPG1 in L. theobromae led to significantly increased and decreased lesion areas, respectively. Further, the high transcript level of LtEPG1 during the infection process supported its virulence function. Polygalacturonase activity of LtEPG1 was substantiated by detecting its ability to degrade pectin. Furthermore, LtEPG1 functioned as microbe-associated molecular patterns during the infection process. Both transient expression of LtEPG1 in planta and infiltration of purified LtEPG1 triggered cell death in Nicotiana benthamiana. Site-directed mutation of LtEPG1 indicated that the enzymatic activity of LtEPG1 is independent from its elicitor activity. A protein kinase, KINβ1, was shown to interact in the yeast two‐hybrid system with LtEPG1. This interaction was further confirmed in vitro using a pull-down assay. Our data indicate that LtEPG1 functions as a polygalacturonase and also serves as an elicitor with two independent mechanisms. Moreover, LtEPG1 may be able to manipulate host immune responses by regulating the KINβ1-mediated signal pathway and consequently promote its own successful infection and symptom development.

Sign in / Sign up

Export Citation Format

Share Document