Secreted subtilisin gene family in Trichophyton rubrum

Jousson, Olivier; Léchenne, Barbara; Bontems, Olympia; Mignon, Bernard; Reichard, Utz; Barblan, Jachen; Quadroni, Manfredo; Monod, Michel

doi:10.1016/j.gene.2004.06.024

Cited by 100 publications

(124 citation statements)

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“…A functional application of the T. rubrum microarray to the analysis of A. benhamiae was estimated, based on the DNA sequence homology between T. rubrum and A. benhamiae, e.g. 91-97 % identity between orthologues of the previously described subtilisin genes SUB1-SUB7 (Jousson et al, 2004b). Samples of A. benhamiae grown in either soy (soyA/B/C) or keratin-soy medium (keratin-soyA/B/C) were compared with probes derived from cells grown in control Sabouraud medium (SabA/B/C).…”

Section: Resultsmentioning

confidence: 99%

“…The expression levels of six A. benhamiae genes were analysed by quantitative reverse-transcription PCR (qRT-PCR). Using PrimerExpress 2.0 software (Applied Biosystems), specific primers and 59-FAM-39BHQ probes were designed for genes encoding A. benhamiae subtilisins Sub3 and Sub6, which have been published previously (Jousson et al, 2004b); the sequence of primer Sub3AbenR is identical to that of primer Sub3TrubR (Zaugg et al, 2009). Sequences of primers and probes for detection of genes encoding McpA and isocitrate lyase as well as the controls encoding ADP-ribosylation factor ADPrf (TrMZC10ACH) and chitin-synthase ChitinS (TrMZG10ACO) were based on T. rubrum sequences (Zaugg et al, 2009).…”

Section: Methodsmentioning

confidence: 99%

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Differential gene expression in the pathogenic dermatophyte Arthroderma benhamiae in vitro versus during infection

et al. 2010

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Although dermatophytes are the most common agents of superficial mycoses in humans and animals, the molecular basis of the pathogenicity of these fungi is largely unknown. In vitro digestion of keratin by dermatophytes is associated with the secretion of multiple proteases, which are assumed to be responsible for their particular specialization to colonize and degrade keratinized host structures during infection. To investigate the role of individual secreted proteases in dermatophytosis, a guinea pig infection model was established for the zoophilic dermatophyte Arthroderma benhamiae, which causes highly inflammatory cutaneous infections in humans and rodents. By use of a cDNA microarray covering approximately 20-25 % of the A. benhamiae genome and containing sequences of at least 23 protease genes, we revealed a distinct in vivo protease gene expression profile in the fungal cells, which was surprisingly different from the pattern elicited during in vitro growth on keratin. Instead of the major in vitroexpressed proteases, others were activated specifically during infection. These enzymes are therefore suggested to fulfil important functions that are not exclusively associated with the degradation of keratin. Most notably, the gene encoding the serine protease subtilisin 6, which is a known major allergen in the related dermatophyte Trichophyton rubrum and putatively linked to host inflammation, was found to be the most strongly upregulated gene during infection. In addition, our approach identified other candidate pathogenicity-related factors in A. benhamiae, such as genes encoding key enzymes of the glyoxylate cycle and an opsin-related protein.Our work provides what we believe to be the first broad-scale gene expression profile in human pathogenic dermatophytes during infection, and points to putative virulence-associated mechanisms that make these micro-organisms the most successful aetiological agents of superficial mycoses.Abbreviations: FDR, false discovery rate; MFS, major facilitator superfamily; PAS, periodic acid Schiff; qRT-PCR, quantitative reverse-transcription PCR.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Differential gene expression in the pathogenic dermatophyte Arthroderma benhamiae in vitro versus during infection

et al. 2010

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“…The hypothesis that one keratinase, alone or with further proteases, decomposes hard keratin, has now been abandoned (Kunert, 1992). Dermatophyte-secreted subtilisins have been shown to be at most 50 % more active towards keratin azure than proteinase K or subtilisin Carlsberg, with the last two enzymes both in the presence of b-mercaptoethanol (Jousson et al, 2004b). Sulphitolysis is so far the sole known dermatophyte mechanism that allows the reduction of disulphide bridges, a bottleneck in the process of degradation of compact keratinized tissues.…”

Section: Discussionmentioning

confidence: 99%

“…Sulphitolysis is so far the sole known dermatophyte mechanism that allows the reduction of disulphide bridges, a bottleneck in the process of degradation of compact keratinized tissues. In contrast, the activities of the dermatophyte-secreted proteases are redundant for the digestion of reduced protein (Jousson et al, 2004b;Monod et al, 2005). Therefore, sulphite transporters, rather than secreted proteases, could be a target of choice to treat dermatophyte infection.…”

Section: Discussionmentioning

confidence: 99%

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Sulphite efflux pumps in Aspergillus fumigatus and dermatophytes

Léchenne

Reichard²,

Zaugg

et al. 2007

Microbiology

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Dermatophytes and other filamentous fungi excrete sulphite as a reducing agent during keratin degradation. In the presence of sulphite, cystine in keratin is directly cleaved to cysteine and Ssulphocysteine, and thereby, reduced proteins become accessible to hydrolysis by a variety of secreted endo-and exoproteases. A gene encoding a sulphite transporter in Aspergillus fumigatus (AfuSSU1), and orthologues in the dermatophytes Trichophyton rubrum and Arthroderma benhamiae (TruSSU1 and AbeSSU1, respectively), were identified by functional expression in Saccharomyces cerevisiae. Like the S. cerevisiae sulphite efflux pump Ssu1p, AfuSsu1p, TruSsu1p and AbeSsu1p belong to the tellurite-resistance/dicarboxylate transporter (TDT) family which includes the Escherichia coli tellurite transporter TehAp and the Schizosaccharomyces pombe malate transporter Mae1p. Seven genes in the A. fumigatus genome encode transporters of the TDT family. However, gene disruption of AfuSSU1 and of the two more closely related paralogues revealed that only AfuSSU1 encodes a sulphite efflux pump. TruSsulp and AbeSsulp are believed to be the first members of the TDT family identified in dermatophytes. The relatively high expression of TruSSU1 and AbeSSU1 in dermatophytes compared to that of AfuSSU1 in A. fumigatus likely reflects a property of dermatophytes which renders these fungi pathogenic. Sulphite transporters could be a new target for antifungal drugs in dermatology, since proteolytic digestion of hard keratin would not be possible without prior reduction of disulphide bridges.

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MALDI / TOF / MS

2004

Encyclopedic Dictionary of Genetics, Genomics and Proteomics

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The Trichophyton rubrum species complex comprises commonly encountered dermatophytic fungi with a worldwide distribution. The members of the complex usually have distinct phenotypes in culture and cause different clinical symptoms, despite high genome similarity. In order to better delimit the species within the complex, molecular, phenotypic, and physiological characteristics were combined to reestablish a natural species concept. Three groups, T. rubrum, T. soudanense, and T. violaceum, could be distinguished based on the sequence of the internal transcribed spacer (ITS) ribosomal DNA barcode gene. On average, strains within each group were similar by colony appearance, microscopy, and physiology, but strains between groups showed significant differences. Trichophyton rubrum strains had higher keratinase activity, whereas T. violaceum strains tended to be more lipophilic; however, none of the phenotypic features were diagnostic. The results of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and amplified fragment length polymorphism (AFLP) were partially consistent with the ITS data but failed to distinguish the species unambiguously. Despite their close similarity, T. violaceum, T. soudanense, and T. rubrum can be regarded as independent species with distinct geographical distributions and clinical predilections. Trichophyton soudanense is pheno-and genotypically intermediate between T. rubrum and T. violaceum. For routine diagnostics, ITS sequencing is recommended.

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Secreted subtilisin gene family in Trichophyton rubrum

Cited by 100 publications

References 33 publications

Differential gene expression in the pathogenic dermatophyte Arthroderma benhamiae in vitro versus during infection

Differential gene expression in the pathogenic dermatophyte Arthroderma benhamiae in vitro versus during infection

Sulphite efflux pumps in Aspergillus fumigatus and dermatophytes

MALDI / TOF / MS

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