scholarly journals The Transcriptional Profile of Trichophyton rubrum Co-Cultured with Human Keratinocytes Shows New Insights about Gene Modulation by Terbinafine

Pathogens ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 274 ◽  
Author(s):  
Monise Fazolin Petrucelli ◽  
Josie Budag Matsuda ◽  
Kamila Peroni ◽  
Pablo Rodrigo Sanches ◽  
Wilson Araújo Silva ◽  
...  
Keyword(s):  
Trichophyton Rubrum ◽  
Human Keratinocytes ◽  
Major Facilitator Superfamily ◽  
Ergosterol Biosynthesis ◽  
Rna Seq ◽  
Host Interaction ◽  
Culture Model ◽  
Superficial Mycosis ◽  
Genes Encoding ◽  
Major Facilitator

The dermatophyte Trichophyton rubrum is the main causative agent of dermatophytoses worldwide. Although a superficial mycosis, its incidence has been increasing especially among diabetic and immunocompromised patients. Terbinafine is commonly used for the treatment of infections caused by dermatophytes. However, cases of resistance of T. rubrum to this allylamine were reported even with the efficacy of this drug. The present study is the first to evaluate the effect of terbinafine using a co-culture model of T. rubrum and human keratinocytes, mimicking a fungus-host interaction, in conjunction with RNA-seq technique. Our data showed the repression of several genes involved in the ergosterol biosynthesis cascade and the induction of genes encoding major facilitator superfamily (MFS)- and ATP-binding cassette superfamily (ABC)-type membrane transporter which may be involved in T. rubrum mechanisms of resistance to this drug. We observed that some genes reported in the scientific literature as candidates of new antifungal targets were also modulated. In addition, we found the modulation of several genes that are hypothetical in T. rubrum but that possess known orthologs in other dermatophytes. Taken together, the results indicate that terbinafine can act on various targets related to the physiology of T. rubrum other than its main target of ergosterol biosynthetic pathway.

scholarly journals Dual RNA-Seq Analysis of Trichophyton rubrum and HaCat Keratinocyte Co-Culture Highlights Important Genes for Fungal-Host Interaction

Genes ◽  
2018 ◽  
Vol 9 (7) ◽  
pp. 362 ◽  
Author(s):  
Monise Petrucelli ◽  
Kamila Peronni ◽  
Pablo Sanches ◽  
Tatiana Komoto ◽  
Josie Matsuda ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Trichophyton Rubrum ◽  
Glyoxylate Cycle ◽  
Human Keratinocytes ◽  
Rna Seq ◽  
Fungal Host ◽  
Host Interaction ◽  
Genes Encoding ◽  
Hacat Keratinocyte

The dermatophyte Trichophyton rubrum is the major fungal pathogen of skin, hair, and nails that uses keratinized substrates as the primary nutrients during infection. Few strategies are available that permit a better understanding of the molecular mechanisms involved in the interaction of T. rubrum with the host because of the limitations of models mimicking this interaction. Dual RNA-seq is a powerful tool to unravel this complex interaction since it enables simultaneous evaluation of the transcriptome of two organisms. Using this technology in an in vitro model of co-culture, this study evaluated the transcriptional profile of genes involved in fungus-host interactions in 24 h. Our data demonstrated the induction of glyoxylate cycle genes, ERG6 and TERG_00916, which encodes a carboxylic acid transporter that may improve the assimilation of nutrients and fungal survival in the host. Furthermore, genes encoding keratinolytic proteases were also induced. In human keratinocytes (HaCat) cells, the SLC11A1, RNASE7, and CSF2 genes were induced and the products of these genes are known to have antimicrobial activity. In addition, the FLG and KRT1 genes involved in the epithelial barrier integrity were inhibited. This analysis showed the modulation of important genes involved in T. rubrum–host interaction, which could represent potential antifungal targets for the treatment of dermatophytoses.

scholarly journals Analysis of saponins detoxification genes in Ilyonectria mors-panacis G3B inducing root rot of Panax notoginseng by RNA-Seq

2021 ◽  
Author(s):  
Guohong Zeng ◽  
Jin Li ◽  
Yuxiu Ma ◽  
Qian Pu ◽  
Tian Xiao ◽  
...  
Keyword(s):  
Root Rot ◽  
Molecular Mechanisms ◽  
Management Strategies ◽  
Panax Notoginseng ◽  
Glycoside Hydrolases ◽  
Rna Seq ◽  
Host Interaction ◽  
Excellent Starting Point ◽  
Starting Point ◽  
Genes Encoding

AbstractSaponins are kinds of antifungal compounds produced by Panax notoginseng to resist invasion by pathogens. Ilyonectria mors-panacis G3B was the dominant pathogen inducing root rot of P. notoginseng, and the abilities to detoxify saponins were the key to infect P. notoginseng successfully. To research the molecular mechanisms of detoxifying saponins in I. mors-panacis G3B, we used high-throughput RNA-Seq to identify 557 and 1519 differential expression genes (DEGs) in I. mors-panacis G3B with saponins treatments for 4H (Hours) and 12H (Hours) compared with no saponins treatments, respectively. Among these DEGs, we found 93 genes which were simultaneously highly expressed in I. mors-panacis G3B with saponins treatments for 4H and 12H, they mainly belong to genes encoding transporters, glycoside hydrolases, oxidation–reduction enzymes, transcription factors and so on. In addition, there were 21 putative PHI (Pathogen–Host Interaction) genes out of those 93 up-regulated genes. In this report, we analyzed virulence-associated genes in I. mors-panacis G3B which may be related to detoxifying saponins to infect P. notoginseng successfully. They provided an excellent starting point for in-depth study on pathogenicity of I. mors-panacis G3B and developed appropriate root rot disease management strategies in the future.

scholarly journals Discovering Novel Bile Protection Systems in Bifidobacterium breve UCC2003 through Functional Genomics

2011 ◽  
Vol 78 (4) ◽  
pp. 1123-1131 ◽  
Author(s):  
Lorena Ruiz ◽  
Aldert Zomer ◽  
Mary O'Connell-Motherway ◽  
Douwe van Sinderen ◽  
Abelardo Margolles
Keyword(s):  
Major Facilitator Superfamily ◽  
Transport Systems ◽  
Content Type ◽  
Bifidobacterium Breve ◽  
Enhanced Resistance ◽  
Protection Systems ◽  
Genes Encoding ◽  
Major Facilitator ◽  
Inducible Genes ◽  
Selection Of

ABSTRACTTolerance of gut commensals to bile salt exposure is an important feature for their survival in and colonization of the intestinal environment. A transcriptomic approach was employed to study the response ofBifidobacterium breveUCC2003 to bile, allowing the identification of a number of bile-induced genes with a range of predicted functions. The potential roles of a selection of these bile-inducible genes in bile protection were analyzed following heterologous expression inLactococcus lactis. Genes encoding three transport systems belonging to the major facilitator superfamily (MFS), Bbr_0838, Bbr_0832, and Bbr_1756, and three ABC-type transporters, Bbr_0406-0407, Bbr_1804-1805, and Bbr_1826-1827, were thus investigated and shown to provide enhanced resistance and survival to bile exposure. This work significantly improves our understanding as to how bifidobacteria respond to and survive bile exposure.

scholarly journals Membrane transporter proteins are involved in Trichophyton rubrum pathogenesis

2009 ◽  
Vol 58 (2) ◽  
pp. 163-168 ◽  
Author(s):  
Fernanda C. A. Maranhão ◽  
Fernanda G. Paião ◽  
Ana Lúcia Fachin ◽  
Nilce M. Martinez-Rossi
Keyword(s):  
Abc Transporter ◽  
Trichophyton Rubrum ◽  
Disease Model ◽  
Initial Step ◽  
Functional Expression ◽  
Infection Process ◽  
Major Facilitator Superfamily ◽  
Gene Encoding ◽  
Major Facilitator

Trichophyton rubrum is a dermatophyte responsible for the majority of human superficial mycoses. The functional expression of proteins important for the initial step and the maintenance of the infection process were identified previously in T. rubrum by subtraction suppression hybridization after growth in the presence of keratin. In this study, sequences similar to genes encoding the multidrug-resistance ATP-binding cassette (ABC) transporter, copper ATPase, the major facilitator superfamily and a permease were isolated, and used in Northern blots to monitor the expression of the genes, which were upregulated in the presence of keratin. A sequence identical to the TruMDR2 gene, encoding an ABC transporter in T. rubrum, was isolated in these experiments, and examination of a T. rubrum ΔTruMDR2 mutant showed a reduction in infecting activity, characterized by low growth on human nails compared with the wild-type strain. The high expression levels of transporter genes by T. rubrum in mimetic infection and the reduction in virulence of the ΔTruMDR2 mutant in a disease model in vitro suggest that transporters are involved in T. rubrum pathogenicity.

scholarly journals A Novel Nitrate/Nitrite Permease in the Marine CyanobacteriumSynechococcus sp. Strain PCC 7002

1999 ◽  
Vol 181 (23) ◽  
pp. 7363-7372 ◽  
Author(s):  
Toshio Sakamoto ◽  
Kaori Inoue-Sakamoto ◽  
Donald A. Bryant
Keyword(s):  
Nitrite Reductase ◽  
Major Facilitator Superfamily ◽  
Wild Type ◽  
Nitrite Uptake ◽  
Genes Encoding ◽  
Major Facilitator ◽  
Mutant Cells ◽  
Nitrate Transporters ◽  
In Cells

ABSTRACT The nrtP and narB genes, encoding nitrate/nitrite permease and nitrate reductase, respectively, were isolated from the marine cyanobacterium Synechococcus sp. strain PCC 7002 and characterized. NrtP is a member of the major facilitator superfamily and is unrelated to the ATP-binding cassette-type nitrate transporters that previously have been described for freshwater strains of cyanobacteria. However, NrtP is similar to the NRT2-type nitrate transporters found in diverse organisms. An nrtP mutant strain consumes nitrate at a 4.5-fold-lower rate than the wild type, and this mutant grew exponentially on a medium containing 12 mM nitrate at a rate approximately 2-fold lower than that of the wild type. The nrtP mutant cells could not consume nitrite as rapidly as the wild type at pH 10, suggesting that NrtP also functions in nitrite uptake. A narB mutant was unable to grow on a medium containing nitrate as a nitrogen source, although this mutant could grow on media containing urea or nitrite with rates similar to those of the wild type. Exogenously added nitrite enhanced the in vivo activity of nitrite reductase in the narBmutant; this suggests that nitrite acts as a positive effector of nitrite reductase. Transcripts of the nrtP andnarB genes were detected in cells grown on nitrate but were not detected in cells grown on urea or ammonia. Transcription of thenrtP and narB genes is probably controlled by the NtcA transcription factor for global nitrogen control. The discovery of a nitrate/nitrite permease in Synechococcussp. strain PCC 7002 suggests that significant differences in nutrient transporters may occur in marine and freshwater cyanobacteria.

scholarly journals Trichophyton rubrum Azole Resistance Mediated by a New ABC Transporter, TruMDR3

2019 ◽  
Vol 63 (11) ◽  
Author(s):  
Michel Monod ◽  
Marc Feuermann ◽  
Karine Salamin ◽  
Marina Fratti ◽  
Maya Makino ◽  
...  
Keyword(s):  
Trichophyton Rubrum ◽  
Heterologous Gene Expression ◽  
Efflux Pumps ◽  
Major Facilitator Superfamily ◽  
Azole Resistance ◽  
Multidrug Efflux ◽  
Content Type ◽  
Milbemycin Oxime ◽  
Major Facilitator ◽  
Mfs Transporters

ABSTRACT The mechanisms of terbinafine resistance in a set of clinical isolates of Trichophyton rubrum have been studied recently. Of these isolates, TIMM20092 also showed reduced sensitivity to azoles. The azole resistance of TIMM20092 could be inhibited by milbemycin oxime, prompting us to examine the potential of T. rubrum to develop resistance through multidrug efflux transporters. The introduction of a T. rubrum cDNA library into Saccharomyces cerevisiae allowed the isolation of one transporter of the major facilitator superfamily (MFS) conferring resistance to azoles (TruMFS1). To identify more azole efflux pumps among 39 ABC and 170 MFS transporters present within the T. rubrum genome, we performed a BLASTp analysis of Aspergillus fumigatus, Candida albicans, and Candida glabrata on transporters that were previously shown to confer azole resistance. The identified candidates were further tested by heterologous gene expression in S. cerevisiae. Four ABC transporters (TruMDR1, TruMDR2, TruMDR3, and TruMDR5) and a second MFS transporter (TruMFS2) proved to be able to operate as azole efflux pumps. Milbemycin oxime inhibited only TruMDR3. Expression analysis showed that both TruMDR3 and TruMDR2 were significantly upregulated in TIMM20092. TruMDR3 transports voriconazole (VRC) and itraconazole (ITC), while TruMDR2 transports only ITC. Disruption of TruMDR3 in TIMM20092 abolished its resistance to VRC and reduced its resistance to ITC. Our study highlights TruMDR3, a newly identified transporter of the ABC family in T. rubrum, which can confer azole resistance if overexpressed. Finally, inhibition of TruMDR3 by milbemycin suggests that milbemycin analogs could be interesting compounds to treat dermatophyte infections in cases of azole resistance.

scholarly journals Transcriptome differences between Cupriavidus necator NH9 grown with 3-chlorobenzoate and that grown with benzoate

2020 ◽  
Author(s):  
Ryota Moriuchi ◽  
Hideo Dohra ◽  
Yu Kanesaki ◽  
Naoto Ogawa
Keyword(s):  
Aromatic Compounds ◽  
Differential Expression Analysis ◽  
Agar Plate ◽  
Enrichment Analysis ◽  
Cupriavidus Necator ◽  
Major Facilitator Superfamily ◽  
Bacterial Degradation ◽  
Genes Encoding ◽  
Chlorinated Aromatic Compounds ◽  
Major Facilitator

Abstract BackgroundAromatic compounds derived from human activities are often released into the environment. Many of them, especially halogenated aromatics, are persistent in nature and pose threats to organisms. Therefore, the microbial degradation of these compounds has been studied intensively. Our laboratory has studied the expression of genes in Cupriavidus necator NH9 involved in the degradation of 3-chlorobenzoate (3-CB), a model compound for studies on bacterial degradation of chlorinated aromatic compounds. In this study aimed at exploring how this bacterium has adapted to the utilization of chlorinated aromatic compounds, we performed RNA-seq analysis of NH9 cells cultured with 3-CB, benzoate (BA), or citric acid (CA). The purpose of these analyses was to identify differentially expressed genes encoding products with various biological functions involved in the degradation of 3-CB and BA.ResultsDifferential expression analysis confirmed strong induction of genes encoding enzymes in degradation pathways of 3-CB and BA, including benABCD (more than 256-fold compared with CA) encoding benzoate 1,2-dioxygenase involved in initial hydroxylation of both 3-CB and BA, and cbnABCD (more than 200-fold compared with BA and CA) encoding enzymes of chlorocatechol ortho-cleavage pathway. Four genes encoding major facilitator superfamily transporters were specifically induced by 3-CB or BA, and one cluster of genes encoding components of the ATP-binding cassette transporter system was significantly induced by 3-CB. Stress response genes encoding chaperones, proteases, the phosphate transporter PstBACS, and superoxide oxidase were upregulated in response to 3-CB and/or BA. Gene Ontology enrichment analysis revealed that genes encoding dioxygenases were upregulated by both 3-CB and BA. Intriguingly, the “cell motility,” “signal transduction,” and “chemotaxis” terms were significantly upregulated by BA compared with 3-CB. Consistent with this, in semi-solid agar plate assays, NH9 cells showed stronger chemotaxis to BA than to 3-CB.ConclusionsOur results showed that the chemotaxis behavior of NH9 differs between 3-CB and BA. We inferred that NH9 has not fully adapted to the utilization of chlorinated benzoate, unlike its analogous aromatic compound BA, in nature.

scholarly journals Transcriptome differences between Cupriavidus necator NH9 grown with 3-chlorobenzoate and that grown with benzoate

2020 ◽  
Author(s):  
Ryota Moriuchi ◽  
Hideo Dohra ◽  
Yu Kanesaki ◽  
Naoto Ogawa
Keyword(s):  
Aromatic Compounds ◽  
Differential Expression Analysis ◽  
Agar Plate ◽  
Enrichment Analysis ◽  
Cupriavidus Necator ◽  
Major Facilitator Superfamily ◽  
Bacterial Degradation ◽  
Genes Encoding ◽  
Chlorinated Aromatic Compounds ◽  
Major Facilitator

Abstract Background Aromatic compounds derived from human activities are often released into the environment. Many of them, especially halogenated aromatics, are persistent in nature and pose threats to organisms. Therefore, the microbial degradation of these compounds has been studied intensively. Our laboratory has studied the expression of genes in Cupriavidus necator NH9 involved in the degradation of 3-chlorobenzoate (3-CB), a model compound for studies on bacterial degradation of chlorinated aromatic compounds. In this study aimed at exploring how this bacterium has adapted to the utilization of chlorinated aromatic compounds, we performed RNA-seq analysis of NH9 cells cultured with 3-CB, benzoate (BA), or citric acid (CA). The purpose of these analyses was to identify differentially expressed genes encoding products with various biological functions involved in the degradation of 3-CB and BA. Results Differential expression analysis confirmed strong induction of genes encoding enzymes in degradation pathways of 3-CB and BA, including benABCD (more than 256-fold compared with CA) encoding benzoate 1,2-dioxygenase involved in initial hydroxylation of both 3-CB and BA, and cbnABCD (more than 200-fold compared with BA and CA) encoding enzymes of chlorocatechol ortho -cleavage pathway. Four genes encoding major facilitator superfamily transporters were specifically induced by 3-CB or BA, and one cluster of genes encoding components of the ATP-binding cassette transporter system was significantly induced by 3-CB. Stress response genes encoding chaperones, proteases, the phosphate transporter PstBACS, and superoxide oxidase were upregulated in response to 3-CB and/or BA. Gene Ontology enrichment analysis revealed that genes encoding dioxygenases were upregulated by both 3-CB and BA. Intriguingly, the “cell motility,” “signal transduction,” and “chemotaxis” terms were significantly upregulated by BA compared with 3-CB. Consistent with this, in semi-solid agar plate assays, NH9 cells showed stronger chemotaxis to BA than to 3-CB. Conclusions Our results showed that the chemotaxis behavior of NH9 differs between 3-CB and BA. We inferred that NH9 has not fully adapted to the utilization of chlorinated benzoate, unlike its analogous aromatic compound BA, in nature.

scholarly journals Identification and Regulation of the N-Acetylglucosamine Utilization Pathway of the Plant Pathogenic Bacterium Xanthomonas campestris pv. campestris

2010 ◽  
Vol 192 (6) ◽  
pp. 1487-1497 ◽  
Author(s):  
Alice Boulanger ◽  
Guillaume Déjean ◽  
Martine Lautier ◽  
Marie Glories ◽  
Claudine Zischek ◽  
...  
Keyword(s):  
Xanthomonas Campestris ◽  
Amino Sugar ◽  
Major Facilitator Superfamily ◽  
Experimental Conditions ◽  
Expression Of Genes ◽  
Wide Range ◽  
Genes Encoding ◽  
Major Facilitator Superfamily Transporter ◽  
Major Facilitator ◽  
Utilization Pathway

ABSTRACT Xanthomonas campestris pv. campestris, the causal agent of black rot disease of brassicas, is known for its ability to catabolize a wide range of plant compounds. This ability is correlated with the presence of specific carbohydrate utilization loci containing TonB-dependent transporters (CUT loci) devoted to scavenging specific carbohydrates. In this study, we demonstrate that there is an X. campestris pv. campestris CUT system involved in the import and catabolism of N-acetylglucosamine (GlcNAc). Expression of genes belonging to this GlcNAc CUT system is under the control of GlcNAc via the LacI family NagR and GntR family NagQ regulators. Analysis of the NagR and NagQ regulons confirmed that GlcNAc utilization involves NagA and NagB-II enzymes responsible for the conversion of GlcNAc-6-phosphate to fructose-6-phosphate. Mutants with mutations in the corresponding genes are sensitive to GlcNAc, as previously reported for Escherichia coli. This GlcNAc sensitivity and analysis of the NagQ and NagR regulons were used to dissect the X. campestris pv. campestris GlcNAc utilization pathway. This analysis revealed specific features, including the fact that uptake of GlcNAc through the inner membrane occurs via a major facilitator superfamily transporter and the fact that this amino sugar is phosphorylated by two proteins belonging to the glucokinase family, NagK-IIA and NagK-IIB. However, NagK-IIA seems to play a more important role in GlcNAc utilization than NagK-IIB under our experimental conditions. The X. campestris pv. campestris GlcNAc NagR regulon includes four genes encoding TonB-dependent active transporters (TBDTs). However, the results of transport experiments suggest that GlcNAc passively diffuses through the bacterial envelope, an observation that calls into question whether GlcNAc is a natural substrate for these TBDTs and consequently is the source of GlcNAc for this nonchitinolytic plant-associated bacterium.

scholarly journals Efflux-Mediated Antibiotic Resistance inAcinetobacterspp.

2010 ◽  
Vol 55 (3) ◽  
pp. 947-953 ◽  
Author(s):  
Sébastien Coyne ◽  
Patrice Courvalin ◽  
Bruno Périchon
Keyword(s):  
Multidrug Resistance ◽  
Acquired Resistance ◽  
Regulatory System ◽  
Major Facilitator Superfamily ◽  
Natural Resistance ◽  
Major Mechanism ◽  
Small Multidrug Resistance ◽  
Genes Encoding ◽  
Resistance Nodulation Division ◽  
Major Facilitator

ABSTRACTAmongAcinetobacterspp.,A. baumanniiis the most frequently implicated in nosocomial infections, in particular in intensive care units. It was initially thought that multidrug resistance (MDR) in this species was due mainly to horizontal acquisition of resistance genes. However, it has recently become obvious that increased expression of chromosomal genes for efflux systems plays a major role in MDR. Among the five superfamilies of pumps, resistance-nodulation-division (RND) systems are the most prevalent in multiply resistantA. baumannii. RND pumps typically exhibit a wide substrate range that can include antibiotics, dyes, biocides, detergents, and antiseptics. Overexpression of AdeABC, secondary to mutations in theadeRSgenes encoding a two-component regulatory system, constitutes a major mechanism of multiresistance inA. baumannii. AdeIJK, intrinsic to this species, is responsible for natural resistance, but since overexpression above a certain threshold is toxic for the host, its contribution to acquired resistance is minimal. The recently described AdeFGH, probably regulated by a LysR-type transcriptional regulator, also confers multidrug resistance when overexpressed. Non-RND efflux systems, such as CraA, AmvA, AbeM, and AbeS, have also been characterized forA. baumannii, as have AdeXYZ and AdeDE for otherAcinetobacterspp. Finally, acquired narrow-spectrum efflux pumps, such as the major facilitator superfamily (MFS) members TetA, TetB, CmlA, and FloR and the small multidrug resistance (SMR) member QacE inAcinetobacterspp., have been detected and are mainly encoded by mobile genetic elements.

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