Cargando…

Development of an efficient gene-targeting system for elucidating infection mechanisms of the fungal pathogen Trichosporon asahii

Trichosporon asahii is a pathogenic fungus that causes severe, deep-seated fungal infections in neutropenic patients. Elucidating the infection mechanisms of T. asahii based on genetic studies requires a specific gene-targeting system. Here, we established an efficient gene-targeting system in a hig...

Descripción completa

Detalles Bibliográficos
Autores principales: Matsumoto, Yasuhiko, Nagamachi, Tae, Yoshikawa, Asami, Yamazaki, Hideki, Yamasaki, Yusuke, Yamada, Tsuyoshi, Sugita, Takashi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8440527/
https://www.ncbi.nlm.nih.gov/pubmed/34521867
http://dx.doi.org/10.1038/s41598-021-97287-3
_version_ 1783752694726393856
author Matsumoto, Yasuhiko
Nagamachi, Tae
Yoshikawa, Asami
Yamazaki, Hideki
Yamasaki, Yusuke
Yamada, Tsuyoshi
Sugita, Takashi
author_facet Matsumoto, Yasuhiko
Nagamachi, Tae
Yoshikawa, Asami
Yamazaki, Hideki
Yamasaki, Yusuke
Yamada, Tsuyoshi
Sugita, Takashi
author_sort Matsumoto, Yasuhiko
collection PubMed
description Trichosporon asahii is a pathogenic fungus that causes severe, deep-seated fungal infections in neutropenic patients. Elucidating the infection mechanisms of T. asahii based on genetic studies requires a specific gene-targeting system. Here, we established an efficient gene-targeting system in a highly pathogenic T. asahii strain identified using the silkworm infection model. By comparing the pathogenicity of T. asahii clinical isolates in a silkworm infection model, T. asahii MPU129 was identified as a highly pathogenic strain. Using an Agrobacterium tumefaciens-mediated gene transfer system, we obtained a T. asahii MPU129 mutant lacking the ku70 gene, which encodes the Ku70 protein involved in the non-homologous end-joining repair of DNA double-strand breaks. The ku70 gene-deficient mutant showed higher gene-targeting efficiency than the wild-type strain for constructing a mutant lacking the cnb1 gene, which encodes the beta-subunit of calcineurin. The cnb1 gene-deficient mutant showed reduced pathogenicity against silkworms compared with the parental strain. These results suggest that an efficient gene-targeting system in a highly pathogenic T. asahii strain is a useful tool for elucidating the molecular mechanisms of T. asahii infection.
format Online
Article
Text
id pubmed-8440527
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-84405272021-09-15 Development of an efficient gene-targeting system for elucidating infection mechanisms of the fungal pathogen Trichosporon asahii Matsumoto, Yasuhiko Nagamachi, Tae Yoshikawa, Asami Yamazaki, Hideki Yamasaki, Yusuke Yamada, Tsuyoshi Sugita, Takashi Sci Rep Article Trichosporon asahii is a pathogenic fungus that causes severe, deep-seated fungal infections in neutropenic patients. Elucidating the infection mechanisms of T. asahii based on genetic studies requires a specific gene-targeting system. Here, we established an efficient gene-targeting system in a highly pathogenic T. asahii strain identified using the silkworm infection model. By comparing the pathogenicity of T. asahii clinical isolates in a silkworm infection model, T. asahii MPU129 was identified as a highly pathogenic strain. Using an Agrobacterium tumefaciens-mediated gene transfer system, we obtained a T. asahii MPU129 mutant lacking the ku70 gene, which encodes the Ku70 protein involved in the non-homologous end-joining repair of DNA double-strand breaks. The ku70 gene-deficient mutant showed higher gene-targeting efficiency than the wild-type strain for constructing a mutant lacking the cnb1 gene, which encodes the beta-subunit of calcineurin. The cnb1 gene-deficient mutant showed reduced pathogenicity against silkworms compared with the parental strain. These results suggest that an efficient gene-targeting system in a highly pathogenic T. asahii strain is a useful tool for elucidating the molecular mechanisms of T. asahii infection. Nature Publishing Group UK 2021-09-14 /pmc/articles/PMC8440527/ /pubmed/34521867 http://dx.doi.org/10.1038/s41598-021-97287-3 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Matsumoto, Yasuhiko
Nagamachi, Tae
Yoshikawa, Asami
Yamazaki, Hideki
Yamasaki, Yusuke
Yamada, Tsuyoshi
Sugita, Takashi
Development of an efficient gene-targeting system for elucidating infection mechanisms of the fungal pathogen Trichosporon asahii
title Development of an efficient gene-targeting system for elucidating infection mechanisms of the fungal pathogen Trichosporon asahii
title_full Development of an efficient gene-targeting system for elucidating infection mechanisms of the fungal pathogen Trichosporon asahii
title_fullStr Development of an efficient gene-targeting system for elucidating infection mechanisms of the fungal pathogen Trichosporon asahii
title_full_unstemmed Development of an efficient gene-targeting system for elucidating infection mechanisms of the fungal pathogen Trichosporon asahii
title_short Development of an efficient gene-targeting system for elucidating infection mechanisms of the fungal pathogen Trichosporon asahii
title_sort development of an efficient gene-targeting system for elucidating infection mechanisms of the fungal pathogen trichosporon asahii
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8440527/
https://www.ncbi.nlm.nih.gov/pubmed/34521867
http://dx.doi.org/10.1038/s41598-021-97287-3
work_keys_str_mv AT matsumotoyasuhiko developmentofanefficientgenetargetingsystemforelucidatinginfectionmechanismsofthefungalpathogentrichosporonasahii
AT nagamachitae developmentofanefficientgenetargetingsystemforelucidatinginfectionmechanismsofthefungalpathogentrichosporonasahii
AT yoshikawaasami developmentofanefficientgenetargetingsystemforelucidatinginfectionmechanismsofthefungalpathogentrichosporonasahii
AT yamazakihideki developmentofanefficientgenetargetingsystemforelucidatinginfectionmechanismsofthefungalpathogentrichosporonasahii
AT yamasakiyusuke developmentofanefficientgenetargetingsystemforelucidatinginfectionmechanismsofthefungalpathogentrichosporonasahii
AT yamadatsuyoshi developmentofanefficientgenetargetingsystemforelucidatinginfectionmechanismsofthefungalpathogentrichosporonasahii
AT sugitatakashi developmentofanefficientgenetargetingsystemforelucidatinginfectionmechanismsofthefungalpathogentrichosporonasahii