<i>Botrytis cinerea</i> PMT4 Is Involved in <i>O</i>-Glycosylation, Cell Wall Organization, Membrane Integrity, and Virulence
Proteins found within the fungal cell wall usually contain both <i>N</i>- and <i>O</i>-oligosaccharides. <i>N</i>-glycosylation is the process where these oligosaccharides (hereinafter: glycans) are attached to asparagine residues, while in <i>O</i>-gl...
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2025-01-01
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| author | Verónica Plaza Alice Pasten Luz A. López-Ramírez Héctor M. Mora-Montes Julia Rubio-Astudillo Evelyn Silva-Moreno Luis Castillo |
| author_facet | Verónica Plaza Alice Pasten Luz A. López-Ramírez Héctor M. Mora-Montes Julia Rubio-Astudillo Evelyn Silva-Moreno Luis Castillo |
| author_sort | Verónica Plaza |
| collection | DOAJ |
| description | Proteins found within the fungal cell wall usually contain both <i>N</i>- and <i>O</i>-oligosaccharides. <i>N</i>-glycosylation is the process where these oligosaccharides (hereinafter: glycans) are attached to asparagine residues, while in <i>O</i>-glycosylation the glycans are covalently bound to serine or threonine residues. The <i>PMT</i> family is grouped into <i>PMT1</i>, <i>PMT2</i>, and <i>PMT4</i> subfamilies. Using bioinformatics analysis within the <i>Botrytis cinerea</i> genome database, an ortholog to <i>Saccharomyces cerevisiae</i> Pmt4 and other fungal species was identified. The aim of this study was to assess the relevance of the <i>bcpmt4</i> gene in <i>B. cinerea</i> glycosylation. For this purpose, the <i>bcpmt4</i> gene was disrupted by homologous recombination in the B05.10 strain using a hygromycin B resistance cassette. Expression of <i>bcpmt4</i> in <i>S. cerevisiae</i> Δ<i>Scpmt4</i> or Δ<i>Scpmt3</i> null mutants restored glycan levels like those observed in the parental strain. The phenotypic analysis showed that Δ<i>bcpmt4</i> null mutants exhibited significant changes in hyphal cell wall composition, including reduced mannan levels and increased amounts of chitin and glucan. Furthermore, the loss of <i>bcpmt4</i> led to decreased glycosylation of glycoproteins in the <i>B. cinerea</i> cell wall. The null mutant lacking <i>PMT4</i> was hypersensitive to a range of cell wall perturbing agents, antifungal drugs, and high hydrostatic pressure. Thus, in addition to their role in glycosylation, the <i>PMT4</i> is required to virulence, biofilm formation, and membrane integrity. This study adds to our knowledge of the role of the <i>B. cinerea bcpmt4</i> gene, which is involved in glycosylation and cell biology, cell wall formation, and antifungal response. |
| format | Article |
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| institution | Kabale University |
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| publishDate | 2025-01-01 |
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| spelling | doaj-art-65b6b86493424e4abc07a9b2dc4a15ae2025-01-24T13:37:26ZengMDPI AGJournal of Fungi2309-608X2025-01-011117110.3390/jof11010071<i>Botrytis cinerea</i> PMT4 Is Involved in <i>O</i>-Glycosylation, Cell Wall Organization, Membrane Integrity, and VirulenceVerónica Plaza0Alice Pasten1Luz A. López-Ramírez2Héctor M. Mora-Montes3Julia Rubio-Astudillo4Evelyn Silva-Moreno5Luis Castillo6Laboratorio de Biología Molecular y Bioquímica, Departamento de Biología, Universidad de La Serena, La Serena 1700000, ChileLaboratorio de Biología Molecular y Bioquímica, Departamento de Biología, Universidad de La Serena, La Serena 1700000, ChileDepartamento de Biología, División de Ciencias Naturales y Exactas, Universidad de Guanajuato, Guanajuato 36050, MexicoDepartamento de Biología, División de Ciencias Naturales y Exactas, Universidad de Guanajuato, Guanajuato 36050, MexicoInstituto de Ciencias Biomédicas, Universidad Autónoma de Chile, Santiago 7500912, ChileGEMA Center for Genomics, Ecology and Environment, Faculty of Interdisciplinary Studies, Universidad Mayor, Santiago 7510041, ChileLaboratorio de Biología Molecular y Bioquímica, Departamento de Biología, Universidad de La Serena, La Serena 1700000, ChileProteins found within the fungal cell wall usually contain both <i>N</i>- and <i>O</i>-oligosaccharides. <i>N</i>-glycosylation is the process where these oligosaccharides (hereinafter: glycans) are attached to asparagine residues, while in <i>O</i>-glycosylation the glycans are covalently bound to serine or threonine residues. The <i>PMT</i> family is grouped into <i>PMT1</i>, <i>PMT2</i>, and <i>PMT4</i> subfamilies. Using bioinformatics analysis within the <i>Botrytis cinerea</i> genome database, an ortholog to <i>Saccharomyces cerevisiae</i> Pmt4 and other fungal species was identified. The aim of this study was to assess the relevance of the <i>bcpmt4</i> gene in <i>B. cinerea</i> glycosylation. For this purpose, the <i>bcpmt4</i> gene was disrupted by homologous recombination in the B05.10 strain using a hygromycin B resistance cassette. Expression of <i>bcpmt4</i> in <i>S. cerevisiae</i> Δ<i>Scpmt4</i> or Δ<i>Scpmt3</i> null mutants restored glycan levels like those observed in the parental strain. The phenotypic analysis showed that Δ<i>bcpmt4</i> null mutants exhibited significant changes in hyphal cell wall composition, including reduced mannan levels and increased amounts of chitin and glucan. Furthermore, the loss of <i>bcpmt4</i> led to decreased glycosylation of glycoproteins in the <i>B. cinerea</i> cell wall. The null mutant lacking <i>PMT4</i> was hypersensitive to a range of cell wall perturbing agents, antifungal drugs, and high hydrostatic pressure. Thus, in addition to their role in glycosylation, the <i>PMT4</i> is required to virulence, biofilm formation, and membrane integrity. This study adds to our knowledge of the role of the <i>B. cinerea bcpmt4</i> gene, which is involved in glycosylation and cell biology, cell wall formation, and antifungal response.https://www.mdpi.com/2309-608X/11/1/71pathogenicityantifungal drugscell membraneglycoprotein |
| spellingShingle | Verónica Plaza Alice Pasten Luz A. López-Ramírez Héctor M. Mora-Montes Julia Rubio-Astudillo Evelyn Silva-Moreno Luis Castillo <i>Botrytis cinerea</i> PMT4 Is Involved in <i>O</i>-Glycosylation, Cell Wall Organization, Membrane Integrity, and Virulence Journal of Fungi pathogenicity antifungal drugs cell membrane glycoprotein |
| title | <i>Botrytis cinerea</i> PMT4 Is Involved in <i>O</i>-Glycosylation, Cell Wall Organization, Membrane Integrity, and Virulence |
| title_full | <i>Botrytis cinerea</i> PMT4 Is Involved in <i>O</i>-Glycosylation, Cell Wall Organization, Membrane Integrity, and Virulence |
| title_fullStr | <i>Botrytis cinerea</i> PMT4 Is Involved in <i>O</i>-Glycosylation, Cell Wall Organization, Membrane Integrity, and Virulence |
| title_full_unstemmed | <i>Botrytis cinerea</i> PMT4 Is Involved in <i>O</i>-Glycosylation, Cell Wall Organization, Membrane Integrity, and Virulence |
| title_short | <i>Botrytis cinerea</i> PMT4 Is Involved in <i>O</i>-Glycosylation, Cell Wall Organization, Membrane Integrity, and Virulence |
| title_sort | i botrytis cinerea i pmt4 is involved in i o i glycosylation cell wall organization membrane integrity and virulence |
| topic | pathogenicity antifungal drugs cell membrane glycoprotein |
| url | https://www.mdpi.com/2309-608X/11/1/71 |
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