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...

Descripción completa

Detalles Bibliográficos
Autores principales: 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
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