Cloning and sequencing of two Candida parapsilosis genes encoding acid proteases

Viragh, Pierre A. de; Sanglard, Dominique; Togni, Giuseppe; Falchetto, Rocco; Monod, Michel

doi:10.1099/00221287-139-2-335

Cited by 63 publications

(35 citation statements)

References 37 publications

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“…(1,20). However, under in vitro conditions, when serum albumin is the major nitrogen source in the growth medium, C. parapsilosis also produces large amounts of one dominant Sap (e.g., Sapp1p, like C. albicans and C. tropicalis, which produce Sap2p and Sapt1p, respectively (7,14,16,20,21,28). Furthermore, the genes encoding these three isoenzymes are coordinately regulated (30).…”

Section: Discussionmentioning

confidence: 99%

Secreted Aspartic Proteinase Family of Candida tropicalis

Zaugg¹,

et al. 2001

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Medically important yeasts of the genus Candida secrete aspartic proteinases (Saps), which are of particular interest as virulence factors. Like Candida albicans, Candida tropicalis secretes in vitro one dominant Sap (Sapt1p) in a medium containing bovine serum albumin (BSA) as the sole source of nitrogen. Using the gene SAPT1 as a probe and under low-stringency hybridization conditions, three new closely related gene sequences, SAPT2 to SAPT4, encoding secreted proteinases were cloned from a C. tropicalis EMBL3 genomic library. All bands identified by Southern blotting of EcoRI-digested C. tropicalis genomic DNA with SAPT1 could be assigned to a specific SAP gene. Therefore, the SAPT gene family of C. tropicalis is likely to contain only four members. Interestingly, the SAPT2 and SAPT3 gene products, Sapt2p and Sapt3p, which have not yet been detected in C. tropicalis cultures in vitro, were produced as active recombinant enzymes with the methylotrophic yeast Pichia pastoris as an expression system. As expected, reverse transcriptase PCR experiments revealed a strong SAPT1 signal with RNA extracted from cells grown in BSA medium. However, a weak signal was obtained with all other SAPT genes under several conditions tested, showing that these SAPT genes could be expressed at a basic level. Together, these experiments suggest that the gene products Sapt2p, Sapt3p, and Sapt4p could be produced under conditions yet to be described in vitro or during infection.

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Section: Discussionmentioning

confidence: 99%

Secreted Aspartic Proteinase Family of Candida tropicalis

Zaugg¹,

et al. 2001

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“…Many of the pathogenic Candida species have been shown to posses SAP genes, including C. dubliniensis (76), C. tropicalis (147,234,254), and C. parapsilosis (53,147), all of which produce active extracellular proteinases in vitro (76,185). C. tropicalis is thought to possess four SAP genes (254), whereas C. parapsilosis possesses at least two SAP genes (53). Little published information is available with regard to the importance of Sap proteins in the virulence of C. dubliniensis.…”

Section: Secreted Aspartyl Proteinases Of Candidamentioning

confidence: 99%

Candida albicansSecreted Aspartyl Proteinases in Virulence and Pathogenesis

Naglik

Challacombe

Hube

2003

Microbiol Mol Biol Rev

1,014

970

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Candida albicans is the most common fungal pathogen of humans and has developed an extensive repertoire of putative virulence mechanisms that allows successful colonization and infection of the host under suitable predisposing conditions. Extracellular proteolytic activity plays a central role in Candida pathogenicity and is produced by a family of 10 secreted aspartyl proteinases (Sap proteins). Although the consequences of proteinase secretion during human infections is not precisely known, in vitro, animal, and human studies have implicated the proteinases in C. albicans virulence in one of the following seven ways: (i) correlation between Sap production in vitro and Candida virulence, (ii) degradation of human proteins and structural analysis in determining Sap substrate specificity, (iii) association of Sap production with other virulence processes of C. albicans, (iv) Sap protein production and Sap immune responses in animal and human infections, (v) SAP gene expression during Candida infections, (vi) modulation of C. albicans virulence by aspartyl proteinase inhibitors, and (vii) the use of SAP-disrupted mutants to analyze C. albicans virulence. Sap proteins fulfill a number of specialized functions during the infective process, which include the simple role of digesting molecules for nutrient acquisition, digesting or distorting host cell membranes to facilitate adhesion and tissue invasion, and digesting cells and molecules of the host immune system to avoid or resist antimicrobial attack by the host. We have critically discussed the data relevant to each of these seven criteria, with specific emphasis on how this proteinase family could contribute to Candida virulence and pathogenesis

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“…have been studied extensively (13). C. albicans, C. tropicalis, C. dubliniensis, C. guilliermondii, and C. parapsilosis possess SAP gene families (7,10,13,16,30). The Saps of C. albicans, C. parapsilosis, C. tropicalis, and C. lusitaniae have been characterized (2,9,12,14,19,22,26), and their inhibitors have been tested as potential antimycotic drugs (for a review, see reference 27).…”

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Simple Method for Screening Candida Species Isolates for the Presence of Secreted Proteinases: a Tool for the Prediction of Successful Inhibitory Treatment

et al. 2003

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The yeasts of the genus Candida are opportunistic pathogens associated with the rising incidence of life-threatening infections in immunocompromised individuals. Secretion of aspartic proteinases has been determined to be one of the virulence factors of the pathogenic Candida species. To analyze the extracellular proteolytic activities of a large number of Candida clinical isolates, we developed a screening system based on a solid medium containing hemoglobin as the sole nitrogen source. The cleavage of hemoglobin by the secreted proteinases results in formation of clearance zones. The visibility of such zones was enhanced by addition of an acid-base indicator. Using this system, we assessed 245 clinical isolates of Candida from patients in the hospital of the Faculty of Medicine, Palacky University, Olomouc, Czech Republic, for the presence of secreted aspartic proteases (Saps). We also used the test plates for rapid semiquantitative testing of Sap inhibitors. Most of the pepstatin analogs affected the formation of the zones of clearance as well as the growth of Candida albicans, C. tropicalis, and C. parapsilosis colonies. By contrast, the human immunodeficiency virus proteinase inhibitors saquinavir, ritonavir, nelfinavir, and indinavir had no effect on the Candida strains tested. These results are in agreement with the inhibition constants obtained for the individual inhibitors with purified Saps. Thus, the plates containing hemoglobin proved to be an appropriate tool for the rapid and reliable assessment of Sap production and inhibition.The yeasts of the genus Candida are opportunistically invasive in individuals whose defense mechanisms are impaired. Pathogenic Candida species cause diseases ranging from superficial mycoses to disseminated and often fatal infections. The individuals at risk include intensive care and postsurgical patients, human immunodeficiency virus (HIV)-infected hosts, patients with hematological malignancies, elderly patients, and premature infants (4,6,18,25,28). Although Candida albicans is the most frequently isolated yeast associated with human infection, changing patterns of the Candida species detected among clinical isolates in the last decade are evident (5,8,24,28). Therefore, rapid and reliable identification of Candida species producing certain virulence factors is important in routine clinical microbiology practice.Virulence attributes of Candida species include adherence to host tissues, morphological changes, and secretion of hydrolases, e.g., phospholipases and proteinases (11). Secreted aspartic proteinases (Saps) of pathogenic Candida spp. have been studied extensively (13). C. albicans, C. tropicalis, C. dubliniensis, C. guilliermondii, and C. parapsilosis possess SAP gene families (7,10,13,16,30). The Saps of C. albicans, C. parapsilosis, C. tropicalis, and C. lusitaniae have been characterized (2,9,12,14,19,22,26), and their inhibitors have been tested as potential antimycotic drugs (for a review, see reference 27). Information on the extracellular proteolysis of o...

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Cloning and sequencing of two Candida parapsilosis genes encoding acid proteases

Cited by 63 publications

References 37 publications

Secreted Aspartic Proteinase Family of Candida tropicalis

Secreted Aspartic Proteinase Family of Candida tropicalis

Candida albicansSecreted Aspartyl Proteinases in Virulence and Pathogenesis

Simple Method for Screening Candida Species Isolates for the Presence of Secreted Proteinases: a Tool for the Prediction of Successful Inhibitory Treatment

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