Molecular cloning and sequencing of the gene encoding an extracellular aspartic proteinase from<i>Aspergillus fumigatus</i>

Reichard, Utz; Monod, Michel; Rüchel, R.

doi:10.1111/j.1574-6968.1995.tb07700.x

Cited by 31 publications

(34 citation statements)

References 26 publications

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“…PrtT regulates transcription of major secreted proteases. As it was evident that the prtT gene product would regulate the expression of extracellular proteolytic activities, we monitored steady-state transcript levels of several selected genes encoding major secreted proteases of A. fumigatus, in particular alp1 (AFUA_84G11800), mep (AFUA_8G07080), pep1 (AFUA_5G13300), sedB/tppA (AFUA_4G03490), dppIV (AFUA_4G09320), and dppV (AFUA_2G09030), which encode a serine alkaline protease (16), a metalloprotease (15), aspergillopepsin (43), a sedolisin (41), and two dipeptidyl-peptidases (2, 3), respectively (Fig. 4).…”

Section: Resultsmentioning

confidence: 99%

A Regulator of Aspergillus fumigatus Extracellular Proteolytic Activity Is Dispensable for Virulence

Bergmann¹,

Hartmann²,

Cairns

et al. 2009

Infect Immun

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Virulence of the fungal pathogen Aspergillus fumigatus is in part based on the saprophytic lifestyle that this mold has evolved. A crucial function for saprophytism resides in secreted proteases that allow assimilation of proteinaceous substrates. The impact of extracellular proteolytic activities on the pathogenesis of aspergillosis, however, remains controversial. In order to address this issue, characterization of a conserved regulatory factor, PrtT, that acts on expression of secreted proteases was pursued. Expression of PrtT appears to be regulated posttranscriptionally, and the existence of an mRNA leader sequence implies translational control via eIF2␣ kinase signaling. Phenotypic classification of a prtT⌬ deletion mutant revealed that expression of several major extracellular proteases is PrtT dependent, resulting in the inability to utilize protein as a nutritional source. Certain genes encoding secreted proteases are not regulated by PrtT. Most strikingly, the deletant strain is not attenuated in virulence when tested in a leukopenic mouse model, which makes a strong case for reconsidering any impact of secreted proteases in pulmonary aspergillosis.

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Section: Resultsmentioning

confidence: 99%

A Regulator of Aspergillus fumigatus Extracellular Proteolytic Activity Is Dispensable for Virulence

Bergmann¹,

Hartmann²,

Cairns

et al. 2009

Infect Immun

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“…Large peptides cannot be used as nutrients. Therefore, SedA and the aspartic protease Pep1 (35) as endoproteases, together with SedB, SedC, and SedD as exoproteases, constitute a set of proteases that can be used by A. fumigatus to degrade proteins at acidic pHs and to generate assimilable nitrogen sources in decomposing organic matter and composts. In addition, as A. fumigatus regularly acidifies its culture supernatant in vitro (32), it is likely to acidify its microenvironment in the living host.…”

Section: Discussionmentioning

confidence: 99%

Sedolisins, a New Class of Secreted Proteases fromAspergillus fumigatuswith Endoprotease or Tripeptidyl-Peptidase Activity at Acidic pHs

Reichard¹,

Léchenne²,

Asif³

et al. 2006

Appl Environ Microbiol

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The secreted proteolytic activity of Aspergillus fumigatus is of potential importance as a virulence factor and in the industrial hydrolysis of protein sources. The A. fumigatus genome contains sequences that could encode a five-member gene family that produces proteases in the sedolisin family (MEROPS S53). Four putative secreted sedolisins with a predicted 17-to 20-amino-acid signal sequence were identified and termed SedA to SedD. SedA produced heterologously in Pichia pastoris was an acidic endoprotease. Heterologously produced SedB, SedC, and SedD were tripeptidyl-peptidases (TPP) with a common specificity for tripeptide-p-nitroanilide substrates at acidic pHs. Purified SedB hydrolyzed the peptide Ala-Pro-Gly-Asp-Arg-Ile-Tyr-Val-HisPro-Phe to Arg-Pro-Gly, Asp-Arg-Ile, and Tyr-Val-His-Pro-Phe, thereby confirming TPP activity of the enzyme. SedB, SedC, and SedD were detected by Western blotting in culture supernatants of A. fumigatus grown in a medium containing hemoglobin as the sole nitrogen source. A degradation product of SedA also was observed. A search for genes encoding sedolisin homologues in other fungal genomes indicates that sedolisin gene families are widespread among filamentous ascomycetes.

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“…(Hube, 1998;Monod et al, 1993), Rhizopus spp. and other aspergilli (Reichard et al, 1995). Proteinases have not been studied intensively in plant-pathogenic fungi (Barkholt, 1987;Clark et al, 1997;Murphy & Walton, 1996;Poussereau et al, 2001b).…”

Section: B Cinerea Cultures Exclusively Contain Secreted Ap Activitymentioning

confidence: 99%

An aspartic proteinase gene family in the filamentous fungus Botrytis cinerea contains members with novel features

et al. 2004

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Botrytis cinerea, an important fungal plant pathogen, secretes aspartic proteinase (AP) activity in axenic cultures. No cysteine, serine or metalloproteinase activity could be detected. Proteinase activity was higher in culture medium containing BSA or wheat germ extract, as compared to minimal medium. A proportion of the enzyme activity remained in the extracellular glucan sheath. AP was also the only type of proteinase activity in fluid obtained from B. cinerea-infected tissue of apple, pepper, tomato and zucchini. Five B. cinerea genes encoding an AP were cloned and denoted Bcap1–5. Features of the encoded proteins are discussed. BcAP1, especially, has novel characteristics. A phylogenetic analysis was performed comprising sequences originating from different kingdoms. BcAP1 and BcAP5 did not cluster in a bootstrap-supported clade. BcAP2 clusters with vacuolar APs. BcAP3 and BcAP4 cluster with secreted APs in a clade that also contains glycosylphosphatidylinositol-anchored proteinases from Saccharomyces cerevisiae and Candida albicans. All five Bcap genes are expressed in liquid cultures. Transcript levels of Bcap1, Bcap2, Bcap3 and Bcap4 are subject to glucose and peptone repression. Transcripts from all five Bcap genes were detected in infected plant tissue, indicating that at least part of the AP activity in planta originates from the pathogen.

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Molecular cloning and sequencing of the gene encoding an extracellular aspartic proteinase fromAspergillus fumigatus

Cited by 31 publications

References 26 publications

A Regulator of Aspergillus fumigatus Extracellular Proteolytic Activity Is Dispensable for Virulence

A Regulator of Aspergillus fumigatus Extracellular Proteolytic Activity Is Dispensable for Virulence

Sedolisins, a New Class of Secreted Proteases fromAspergillus fumigatuswith Endoprotease or Tripeptidyl-Peptidase Activity at Acidic pHs

An aspartic proteinase gene family in the filamentous fungus Botrytis cinerea contains members with novel features

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