Cyclosporine Biosynthesis in <italic toggle="yes">Tolypocladium inflatum</italic> Benefits Fungal Adaptation to the Environment

ABSTRACT The cycloundecapeptide cyclosporin A (CsA) was first isolated from the insect-pathogenic fungus Tolypocladium inflatum for its antifungal activity and later developed as an immunosuppressant drug. However, the full biosynthetic mechanism of CsA remains unknown and has puzzled researchers fo...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Xiuqing Yang, Peng Feng, Ying Yin, Kathryn Bushley, Joseph W. Spatafora, Chengshu Wang
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	cyclosporine Tolypocladium inflatum antifungal activity biosynthetic pathway virulence Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/a2fe4496fd814f0592509202ec8e0ef0
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:a2fe4496fd814f0592509202ec8e0ef0
record_format	dspace
spelling	oai:doaj.org-article:a2fe4496fd814f0592509202ec8e0ef02021-11-15T15:58:20ZCyclosporine Biosynthesis in <italic toggle="yes">Tolypocladium inflatum</italic> Benefits Fungal Adaptation to the Environment10.1128/mBio.01211-182150-7511https://doaj.org/article/a2fe4496fd814f0592509202ec8e0ef02018-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01211-18https://doaj.org/toc/2150-7511ABSTRACT The cycloundecapeptide cyclosporin A (CsA) was first isolated from the insect-pathogenic fungus Tolypocladium inflatum for its antifungal activity and later developed as an immunosuppressant drug. However, the full biosynthetic mechanism of CsA remains unknown and has puzzled researchers for decades. In this study, the biosynthetic gene cluster is suggested to include 12 genes encoding enzymes, including the nonribosomal peptide synthetase (NRPS) (SimA) responsible for assembling the 11 amino acid substrates of cyclosporine and a polyketide synthase (PKS) (SimG) to mediate the production of the unusual amino acid (4R)-4-[(E)-2-butenyl]-4-methyl-l-threonine (Bmt). Individual deletion of 10 genes, isolation of intermediates, and substrate feeding experiments show that Bmt is biosynthesized by three enzymes, including SimG, SimI, and SimJ. The substrate d-alanine is catalyzed from l-alanine by alanine racemase SimB. Gene cluster transcription is regulated by a putative basic leucine zipper (bZIP)-type protein encoded by the cluster gene SimL. We also found that the cluster cyclophilin (SimC) and transporter (SimD) genes contribute to the tolerance of CsA in the CsA-producing fungus. We also found that cyclosporine production could enable the fungus to outcompete other fungi during cocultivation tests. Deletion of the CsA biosynthetic genes also impaired fungal virulence against insect hosts. Taking all the data together, in addition to proposing a biosynthetic pathway of cyclosporines, the results of this study suggest that CsA produced by this fungus might play important ecological roles in fungal environment interactions. IMPORTANCE The cyclopeptide cyclosporin A was first isolated from the filamentous fungus Tolypocladium inflatum showing antifungal activity and was later developed as an immunosuppressant drug. We report the biosynthetic mechanism of cyclosporines that are mediated by a cluster of genes encoding NRPS and PKS controlled by a bZIP-type transcriptional regulator. The two unusual amino acids Bmt and d-Ala are produced by the PKS pathway and alanine racemase, respectively. The cyclophilin and transporter genes jointly contribute to fungal self-protection against cyclosporines. Cyclosporine confers on T. inflatum the abilities to outcompete other fungi in competitive interactions and to facilitate fungal infection of insect hosts, which therefore benefits fungal adaptations to different environments.Xiuqing YangPeng FengYing YinKathryn BushleyJoseph W. SpataforaChengshu WangAmerican Society for MicrobiologyarticlecyclosporineTolypocladium inflatumantifungal activitybiosynthetic pathwayvirulenceMicrobiologyQR1-502ENmBio, Vol 9, Iss 5 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	cyclosporine Tolypocladium inflatum antifungal activity biosynthetic pathway virulence Microbiology QR1-502
spellingShingle	cyclosporine Tolypocladium inflatum antifungal activity biosynthetic pathway virulence Microbiology QR1-502 Xiuqing Yang Peng Feng Ying Yin Kathryn Bushley Joseph W. Spatafora Chengshu Wang Cyclosporine Biosynthesis in <italic toggle="yes">Tolypocladium inflatum</italic> Benefits Fungal Adaptation to the Environment
description	ABSTRACT The cycloundecapeptide cyclosporin A (CsA) was first isolated from the insect-pathogenic fungus Tolypocladium inflatum for its antifungal activity and later developed as an immunosuppressant drug. However, the full biosynthetic mechanism of CsA remains unknown and has puzzled researchers for decades. In this study, the biosynthetic gene cluster is suggested to include 12 genes encoding enzymes, including the nonribosomal peptide synthetase (NRPS) (SimA) responsible for assembling the 11 amino acid substrates of cyclosporine and a polyketide synthase (PKS) (SimG) to mediate the production of the unusual amino acid (4R)-4-[(E)-2-butenyl]-4-methyl-l-threonine (Bmt). Individual deletion of 10 genes, isolation of intermediates, and substrate feeding experiments show that Bmt is biosynthesized by three enzymes, including SimG, SimI, and SimJ. The substrate d-alanine is catalyzed from l-alanine by alanine racemase SimB. Gene cluster transcription is regulated by a putative basic leucine zipper (bZIP)-type protein encoded by the cluster gene SimL. We also found that the cluster cyclophilin (SimC) and transporter (SimD) genes contribute to the tolerance of CsA in the CsA-producing fungus. We also found that cyclosporine production could enable the fungus to outcompete other fungi during cocultivation tests. Deletion of the CsA biosynthetic genes also impaired fungal virulence against insect hosts. Taking all the data together, in addition to proposing a biosynthetic pathway of cyclosporines, the results of this study suggest that CsA produced by this fungus might play important ecological roles in fungal environment interactions. IMPORTANCE The cyclopeptide cyclosporin A was first isolated from the filamentous fungus Tolypocladium inflatum showing antifungal activity and was later developed as an immunosuppressant drug. We report the biosynthetic mechanism of cyclosporines that are mediated by a cluster of genes encoding NRPS and PKS controlled by a bZIP-type transcriptional regulator. The two unusual amino acids Bmt and d-Ala are produced by the PKS pathway and alanine racemase, respectively. The cyclophilin and transporter genes jointly contribute to fungal self-protection against cyclosporines. Cyclosporine confers on T. inflatum the abilities to outcompete other fungi in competitive interactions and to facilitate fungal infection of insect hosts, which therefore benefits fungal adaptations to different environments.
format	article
author	Xiuqing Yang Peng Feng Ying Yin Kathryn Bushley Joseph W. Spatafora Chengshu Wang
author_facet	Xiuqing Yang Peng Feng Ying Yin Kathryn Bushley Joseph W. Spatafora Chengshu Wang
author_sort	Xiuqing Yang
title	Cyclosporine Biosynthesis in <italic toggle="yes">Tolypocladium inflatum</italic> Benefits Fungal Adaptation to the Environment
title_short	Cyclosporine Biosynthesis in <italic toggle="yes">Tolypocladium inflatum</italic> Benefits Fungal Adaptation to the Environment
title_full	Cyclosporine Biosynthesis in <italic toggle="yes">Tolypocladium inflatum</italic> Benefits Fungal Adaptation to the Environment
title_fullStr	Cyclosporine Biosynthesis in <italic toggle="yes">Tolypocladium inflatum</italic> Benefits Fungal Adaptation to the Environment
title_full_unstemmed	Cyclosporine Biosynthesis in <italic toggle="yes">Tolypocladium inflatum</italic> Benefits Fungal Adaptation to the Environment
title_sort	cyclosporine biosynthesis in <italic toggle="yes">tolypocladium inflatum</italic> benefits fungal adaptation to the environment
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/a2fe4496fd814f0592509202ec8e0ef0
work_keys_str_mv	AT xiuqingyang cyclosporinebiosynthesisinitalictoggleyestolypocladiuminflatumitalicbenefitsfungaladaptationtotheenvironment AT pengfeng cyclosporinebiosynthesisinitalictoggleyestolypocladiuminflatumitalicbenefitsfungaladaptationtotheenvironment AT yingyin cyclosporinebiosynthesisinitalictoggleyestolypocladiuminflatumitalicbenefitsfungaladaptationtotheenvironment AT kathrynbushley cyclosporinebiosynthesisinitalictoggleyestolypocladiuminflatumitalicbenefitsfungaladaptationtotheenvironment AT josephwspatafora cyclosporinebiosynthesisinitalictoggleyestolypocladiuminflatumitalicbenefitsfungaladaptationtotheenvironment AT chengshuwang cyclosporinebiosynthesisinitalictoggleyestolypocladiuminflatumitalicbenefitsfungaladaptationtotheenvironment
_version_	1718427042929704960

Cyclosporine Biosynthesis in <italic toggle="yes">Tolypocladium inflatum</italic> Benefits Fungal Adaptation to the Environment

Ejemplares similares