<i>Ustilago may dis</i> KP6 Killer Toxin: Structure, Expression in <i>Saccharomyces cerevisiae,</i> and Relationship to Other Cellular Toxins

Tao, Jianshi; Ginsberg, I; Banerjee, Nanditta; Held, William A.; Koltin, Y.; Bruenn, Jeremy A.

doi:10.1128/mcb.10.4.1373-1381.1990

Cited by 3 publications

(1 citation statement)

References 48 publications

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“…On the other hand, a PAF orthologous gene is known as antifungal protein from ascomycetes and is identified in the genome of Giberella zeae and named as G. zeae antimicrobial protein A. Microorganisms especially fungi can produce secondary metabolite that have potential to inhibit the growth of the pathogens. Saccharomyces cerevisiae produce toxins and are named killer toxins (Tao et al, 1990). The study about killer toxin from Ustilago maydis has an effective killer activity against Brettanomyces bruxellensis (Santos, Navascues, Bravo, & Marquina, 2011).…”

Section: A B Cmentioning

confidence: 99%

Purification and Identification of an Antifungal Protein from an Isolated Fungus with Antagonism to Colletotrichum gloeosporioides MC9

2022

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Colletotrichum gloeosporioides is the cause of anthracnose disease on mango. This disease becomes more damaging because it economically affects the harvested fruit during the postharvest season. In this research, eight isolates are isolated from the soil of a mango plantation. One of the isolates shows antifungal activity against C. gloeosporioides MC9. This isolate is identified as Penicillium citrinum isolate S1 based on the phylogenetic analysis of ribosomal rRNA sequence. From the culture of this isolate, extracellular filtrates are collected and evaluated for their antifungal activity. The mycelial growth of C. gloeosporioides is significantly inhibited by the culture supernatant of P. citrinum isolate S1. The culture filtrate is used to purify the antifungal protein using ammonium sulfate and ultrafiltration methods. Results show that the antifungal protein was estimated at around 40 kDa molecular weight when separated on a 10% Sodium dodecyl sulfate-polyacrylamide gel. After nine days of incubation, this antifungal protein's inhibition effect with a concentration of 0.94 mg/ml remained 63.6% against C. gloeosporioides. The LCMS result showed that the antifungal protein belongs to the L-asparaginase superfamily. Based on this result, the antifungal protein produced by P. citrinum S1 has the potential to control mango anthracnose disease caused by C. gloeosporioides.

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Section: A B Cmentioning

confidence: 99%

Purification and Identification of an Antifungal Protein from an Isolated Fungus with Antagonism to Colletotrichum gloeosporioides MC9

2022

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Active Internalization of the Penicillium chrysogenum Antifungal Protein PAF in Sensitive Aspergilli

Oberparleiter¹,

Kaiserer²,

Haas³

et al. 2003

Antimicrob Agents Chemother

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The Penicillium chrysogenum antifungal protein PAF inhibits the growth of various filamentous fungi. In this study, PAF was found to localize to the cytoplasm of sensitive aspergilli by indirect immunofluorescence staining. The internalization process required active metabolism and ATP and was prevented by latrunculin B, suggesting an endocytotic mechanism.Antimicrobial proteins are produced by many organisms, including humans, amphibians, arthropods, plants, and fungi (6,11,13,32,37). The Penicillium antifungal protein PAF is abundantly secreted into the supernatant of the ␤-lactam-producing mold Penicillium chrysogenum (22). This small, basic, and cysteine-rich protein specifically inhibits the growth of numerous filamentous fungi (16). Although its primary amino acid structure resembles that of antifungal proteins isolated from other molds, e.g., Aspergillus giganteus (AFP), A. niger (ANAFP), and P. nalgiovense (NAF), significant differences exist in their species specificity (12,20,38). Knowledge of the bases of this selectivity is essential for determining strategies by which to overcome the resistance of important pathogens and to design new antifungal agents. Most data on the possible mechanism of action derive from studies on plant antifungal proteins and AFP which support the hypothesis of an interaction with the plasma membrane that would result in its permeabilization (18,34,35). So far, experiments have addressed such an interaction only indirectly. Therefore, we were interested in elucidating the site of action of the antifungal protein PAF from P. chrysogenum in order to gain better insight into putative target structures that might play a role in the activity of the protein. Here we report the localization of PAF in the sensitive molds A. nidulans, A. fumigatus, and A. niger and give evidence for an active transport of the antifungal protein into affected hyphae.Purification of PAF and generation of polyclonal antiserum. PAF was purified as described previously (16), and polyclonal antibodies against PAF were raised in rabbits as reported elsewhere (26). To determine antibody specificity and the optimal antibody concentration, 25 to 200 ng of PAF was immobilized on Biotrace-NT 0.45-m nitrocellulose membrane strips (PALL Corp., Ann Arbor, Mich.) and detected with various dilutions of rabbit anti-PAF serum and alkaline phosphataseconjugated goat anti-rabbit immunoglobulin G (IgG; 1:10,000; Sigma, Vienna, Austria) in accordance with the protocol previously described (21). The polyclonal antiserum reacted specifically with PAF, and no signals were obtained with the control serum that was collected before the first injection (data not shown).Localization of PAF in aspergilli. Fungi were grown overnight on glass coverslips at 30°C in complete medium CM (16) inoculated with 10 6 conidia/ml. The samples were stained in accordance with the protocol of Fischer et al. (9). In brief, prior to fixation, hyphae were treated with 10 to 50 g of PAF/ml in CM for 90 min at room temperature. The samples were incubat...

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Chitinases Are Essential for Cell Separation in Ustilago maydis

et al. 2015

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Chitin is an essential component of the fungal cell wall, providing rigidity and stability. Its degradation is mediated by chitinases and supposedly ensures the dynamic plasticity of the cell wall during growth and morphogenesis. Hence, chitinases should be particularly important for fungi with dramatic morphological changes, such as Ustilago maydis. This smut fungus switches from yeast to filamentous growth for plant infection, proliferates as a mycelium in planta, and forms teliospores for spreading. Here, we investigate the contribution of its four chitinolytic enzymes to the different morphological changes during the complete life cycle in a comprehensive study of deletion strains combined with biochemical and cell biological approaches. Interestingly, two chitinases act redundantly in cell separation during yeast growth. They mediate the degradation of remnant chitin in the fragmentation zone between mother and daughter cell. In contrast, even the complete lack of chitinolytic activity does not affect formation of the infectious filament, infection, biotrophic growth, or teliospore germination. Thus, unexpectedly we can exclude a major role for chitinolytic enzymes in morphogenesis or pathogenicity of U. maydis. Nevertheless, redundant activity of even two chitinases is essential for cell separation during saprophytic growth, possibly to improve nutrient access or spreading of yeast cells by wind or rain.

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Ustilago may dis KP6 Killer Toxin: Structure, Expression in Saccharomyces cerevisiae, and Relationship to Other Cellular Toxins

Cited by 3 publications

References 48 publications

Purification and Identification of an Antifungal Protein from an Isolated Fungus with Antagonism to Colletotrichum gloeosporioides MC9

Purification and Identification of an Antifungal Protein from an Isolated Fungus with Antagonism to Colletotrichum gloeosporioides MC9

Active Internalization of the Penicillium chrysogenum Antifungal Protein PAF in Sensitive Aspergilli

Chitinases Are Essential for Cell Separation in Ustilago maydis

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