Cargando…
Functional Analysis of the α-1,3-Glucan Synthase Genes agsA and agsB in Aspergillus nidulans: AgsB Is the Major α-1,3-Glucan Synthase in This Fungus
Although α-1,3-glucan is one of the major cell wall polysaccharides in filamentous fungi, the physiological roles of α-1,3-glucan remain unclear. The model fungus Aspergillus nidulans possesses two α-1,3-glucan synthase (AGS) genes, agsA and agsB. For functional analysis of these genes, we construct...
Autores principales: | , , , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3554689/ https://www.ncbi.nlm.nih.gov/pubmed/23365684 http://dx.doi.org/10.1371/journal.pone.0054893 |
_version_ | 1782256953626460160 |
---|---|
author | Yoshimi, Akira Sano, Motoaki Inaba, Azusa Kokubun, Yuko Fujioka, Tomonori Mizutani, Osamu Hagiwara, Daisuke Fujikawa, Takashi Nishimura, Marie Yano, Shigekazu Kasahara, Shin Shimizu, Kiminori Yamaguchi, Masashi Kawakami, Kazuyoshi Abe, Keietsu |
author_facet | Yoshimi, Akira Sano, Motoaki Inaba, Azusa Kokubun, Yuko Fujioka, Tomonori Mizutani, Osamu Hagiwara, Daisuke Fujikawa, Takashi Nishimura, Marie Yano, Shigekazu Kasahara, Shin Shimizu, Kiminori Yamaguchi, Masashi Kawakami, Kazuyoshi Abe, Keietsu |
author_sort | Yoshimi, Akira |
collection | PubMed |
description | Although α-1,3-glucan is one of the major cell wall polysaccharides in filamentous fungi, the physiological roles of α-1,3-glucan remain unclear. The model fungus Aspergillus nidulans possesses two α-1,3-glucan synthase (AGS) genes, agsA and agsB. For functional analysis of these genes, we constructed several mutant strains in A. nidulans: agsA disruption, agsB disruption, and double-disruption strains. We also constructed several CagsB strains in which agsB expression was controlled by the inducible alcA promoter, with or without the agsA-disrupting mutation. The agsA disruption strains did not show markedly different phenotypes from those of the wild-type strain. The agsB disruption strains formed dispersed hyphal cells under liquid culture conditions, regardless of the agsA genetic background. Dispersed hyphal cells were also observed in liquid culture of the CagsB strains when agsB expression was repressed, whereas these strains grew normally in plate culture even under the agsB-repressed conditions. Fractionation of the cell wall based on the alkali solubility of its components, quantification of sugars, and (13)C-NMR spectroscopic analysis revealed that α-1,3-glucan was the main component of the alkali-soluble fraction in the wild-type and agsA disruption strains, but almost no α-1,3-glucan was found in the alkali-soluble fraction derived from either the agsB disruption strain or the CagsB strain under the agsB-repressed conditions, regardless of the agsA genetic background. Taken together, our data demonstrate that the two AGS genes are dispensable in A. nidulans, but that AgsB is required for normal growth characteristics under liquid culture conditions and is the major AGS in this species. |
format | Online Article Text |
id | pubmed-3554689 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-35546892013-01-30 Functional Analysis of the α-1,3-Glucan Synthase Genes agsA and agsB in Aspergillus nidulans: AgsB Is the Major α-1,3-Glucan Synthase in This Fungus Yoshimi, Akira Sano, Motoaki Inaba, Azusa Kokubun, Yuko Fujioka, Tomonori Mizutani, Osamu Hagiwara, Daisuke Fujikawa, Takashi Nishimura, Marie Yano, Shigekazu Kasahara, Shin Shimizu, Kiminori Yamaguchi, Masashi Kawakami, Kazuyoshi Abe, Keietsu PLoS One Research Article Although α-1,3-glucan is one of the major cell wall polysaccharides in filamentous fungi, the physiological roles of α-1,3-glucan remain unclear. The model fungus Aspergillus nidulans possesses two α-1,3-glucan synthase (AGS) genes, agsA and agsB. For functional analysis of these genes, we constructed several mutant strains in A. nidulans: agsA disruption, agsB disruption, and double-disruption strains. We also constructed several CagsB strains in which agsB expression was controlled by the inducible alcA promoter, with or without the agsA-disrupting mutation. The agsA disruption strains did not show markedly different phenotypes from those of the wild-type strain. The agsB disruption strains formed dispersed hyphal cells under liquid culture conditions, regardless of the agsA genetic background. Dispersed hyphal cells were also observed in liquid culture of the CagsB strains when agsB expression was repressed, whereas these strains grew normally in plate culture even under the agsB-repressed conditions. Fractionation of the cell wall based on the alkali solubility of its components, quantification of sugars, and (13)C-NMR spectroscopic analysis revealed that α-1,3-glucan was the main component of the alkali-soluble fraction in the wild-type and agsA disruption strains, but almost no α-1,3-glucan was found in the alkali-soluble fraction derived from either the agsB disruption strain or the CagsB strain under the agsB-repressed conditions, regardless of the agsA genetic background. Taken together, our data demonstrate that the two AGS genes are dispensable in A. nidulans, but that AgsB is required for normal growth characteristics under liquid culture conditions and is the major AGS in this species. Public Library of Science 2013-01-24 /pmc/articles/PMC3554689/ /pubmed/23365684 http://dx.doi.org/10.1371/journal.pone.0054893 Text en © 2013 Yoshimi et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Yoshimi, Akira Sano, Motoaki Inaba, Azusa Kokubun, Yuko Fujioka, Tomonori Mizutani, Osamu Hagiwara, Daisuke Fujikawa, Takashi Nishimura, Marie Yano, Shigekazu Kasahara, Shin Shimizu, Kiminori Yamaguchi, Masashi Kawakami, Kazuyoshi Abe, Keietsu Functional Analysis of the α-1,3-Glucan Synthase Genes agsA and agsB in Aspergillus nidulans: AgsB Is the Major α-1,3-Glucan Synthase in This Fungus |
title | Functional Analysis of the α-1,3-Glucan Synthase Genes agsA and agsB in Aspergillus nidulans: AgsB Is the Major α-1,3-Glucan Synthase in This Fungus |
title_full | Functional Analysis of the α-1,3-Glucan Synthase Genes agsA and agsB in Aspergillus nidulans: AgsB Is the Major α-1,3-Glucan Synthase in This Fungus |
title_fullStr | Functional Analysis of the α-1,3-Glucan Synthase Genes agsA and agsB in Aspergillus nidulans: AgsB Is the Major α-1,3-Glucan Synthase in This Fungus |
title_full_unstemmed | Functional Analysis of the α-1,3-Glucan Synthase Genes agsA and agsB in Aspergillus nidulans: AgsB Is the Major α-1,3-Glucan Synthase in This Fungus |
title_short | Functional Analysis of the α-1,3-Glucan Synthase Genes agsA and agsB in Aspergillus nidulans: AgsB Is the Major α-1,3-Glucan Synthase in This Fungus |
title_sort | functional analysis of the α-1,3-glucan synthase genes agsa and agsb in aspergillus nidulans: agsb is the major α-1,3-glucan synthase in this fungus |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3554689/ https://www.ncbi.nlm.nih.gov/pubmed/23365684 http://dx.doi.org/10.1371/journal.pone.0054893 |
work_keys_str_mv | AT yoshimiakira functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT sanomotoaki functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT inabaazusa functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT kokubunyuko functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT fujiokatomonori functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT mizutaniosamu functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT hagiwaradaisuke functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT fujikawatakashi functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT nishimuramarie functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT yanoshigekazu functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT kasaharashin functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT shimizukiminori functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT yamaguchimasashi functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT kawakamikazuyoshi functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus AT abekeietsu functionalanalysisofthea13glucansynthasegenesagsaandagsbinaspergillusnidulansagsbisthemajora13glucansynthaseinthisfungus |