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Identification and Characterization of a Novel Diterpene Gene Cluster in Aspergillus nidulans
Fungal secondary metabolites are a rich source of medically useful compounds due to their pharmaceutical and toxic properties. Sequencing of fungal genomes has revealed numerous secondary metabolite gene clusters, yet products of many of these biosynthetic pathways are unknown since the expression o...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3323652/ https://www.ncbi.nlm.nih.gov/pubmed/22506079 http://dx.doi.org/10.1371/journal.pone.0035450 |
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author | Bromann, Kirsi Toivari, Mervi Viljanen, Kaarina Vuoristo, Anu Ruohonen, Laura Nakari-Setälä, Tiina |
author_facet | Bromann, Kirsi Toivari, Mervi Viljanen, Kaarina Vuoristo, Anu Ruohonen, Laura Nakari-Setälä, Tiina |
author_sort | Bromann, Kirsi |
collection | PubMed |
description | Fungal secondary metabolites are a rich source of medically useful compounds due to their pharmaceutical and toxic properties. Sequencing of fungal genomes has revealed numerous secondary metabolite gene clusters, yet products of many of these biosynthetic pathways are unknown since the expression of the clustered genes usually remains silent in normal laboratory conditions. Therefore, to discover new metabolites, it is important to find ways to induce the expression of genes in these otherwise silent biosynthetic clusters. We discovered a novel secondary metabolite in Aspergillus nidulans by predicting a biosynthetic gene cluster with genomic mining. A Zn(II)(2)Cys(6)–type transcription factor, PbcR, was identified, and its role as a pathway-specific activator for the predicted gene cluster was demonstrated. Overexpression of pbcR upregulated the transcription of seven genes in the identified cluster and led to the production of a diterpene compound, which was characterized with GC/MS as ent-pimara-8(14),15-diene. A change in morphology was also observed in the strains overexpressing pbcR. The activation of a cryptic gene cluster by overexpression of its putative Zn(II)(2)Cys(6)–type transcription factor led to discovery of a novel secondary metabolite in Aspergillus nidulans. Quantitative real-time PCR and DNA array analysis allowed us to predict the borders of the biosynthetic gene cluster. Furthermore, we identified a novel fungal pimaradiene cyclase gene as well as genes encoding 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase and a geranylgeranyl pyrophosphate (GGPP) synthase. None of these genes have been previously implicated in the biosynthesis of terpenes in Aspergillus nidulans. These results identify the first Aspergillus nidulans diterpene gene cluster and suggest a biosynthetic pathway for ent-pimara-8(14),15-diene. |
format | Online Article Text |
id | pubmed-3323652 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-33236522012-04-13 Identification and Characterization of a Novel Diterpene Gene Cluster in Aspergillus nidulans Bromann, Kirsi Toivari, Mervi Viljanen, Kaarina Vuoristo, Anu Ruohonen, Laura Nakari-Setälä, Tiina PLoS One Research Article Fungal secondary metabolites are a rich source of medically useful compounds due to their pharmaceutical and toxic properties. Sequencing of fungal genomes has revealed numerous secondary metabolite gene clusters, yet products of many of these biosynthetic pathways are unknown since the expression of the clustered genes usually remains silent in normal laboratory conditions. Therefore, to discover new metabolites, it is important to find ways to induce the expression of genes in these otherwise silent biosynthetic clusters. We discovered a novel secondary metabolite in Aspergillus nidulans by predicting a biosynthetic gene cluster with genomic mining. A Zn(II)(2)Cys(6)–type transcription factor, PbcR, was identified, and its role as a pathway-specific activator for the predicted gene cluster was demonstrated. Overexpression of pbcR upregulated the transcription of seven genes in the identified cluster and led to the production of a diterpene compound, which was characterized with GC/MS as ent-pimara-8(14),15-diene. A change in morphology was also observed in the strains overexpressing pbcR. The activation of a cryptic gene cluster by overexpression of its putative Zn(II)(2)Cys(6)–type transcription factor led to discovery of a novel secondary metabolite in Aspergillus nidulans. Quantitative real-time PCR and DNA array analysis allowed us to predict the borders of the biosynthetic gene cluster. Furthermore, we identified a novel fungal pimaradiene cyclase gene as well as genes encoding 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase and a geranylgeranyl pyrophosphate (GGPP) synthase. None of these genes have been previously implicated in the biosynthesis of terpenes in Aspergillus nidulans. These results identify the first Aspergillus nidulans diterpene gene cluster and suggest a biosynthetic pathway for ent-pimara-8(14),15-diene. Public Library of Science 2012-04-10 /pmc/articles/PMC3323652/ /pubmed/22506079 http://dx.doi.org/10.1371/journal.pone.0035450 Text en Bromann 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 Bromann, Kirsi Toivari, Mervi Viljanen, Kaarina Vuoristo, Anu Ruohonen, Laura Nakari-Setälä, Tiina Identification and Characterization of a Novel Diterpene Gene Cluster in Aspergillus nidulans |
title | Identification and Characterization of a Novel Diterpene Gene Cluster in Aspergillus nidulans
|
title_full | Identification and Characterization of a Novel Diterpene Gene Cluster in Aspergillus nidulans
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title_fullStr | Identification and Characterization of a Novel Diterpene Gene Cluster in Aspergillus nidulans
|
title_full_unstemmed | Identification and Characterization of a Novel Diterpene Gene Cluster in Aspergillus nidulans
|
title_short | Identification and Characterization of a Novel Diterpene Gene Cluster in Aspergillus nidulans
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title_sort | identification and characterization of a novel diterpene gene cluster in aspergillus nidulans |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3323652/ https://www.ncbi.nlm.nih.gov/pubmed/22506079 http://dx.doi.org/10.1371/journal.pone.0035450 |
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