Genome-Wide Chromatin Immunoprecipitation Sequencing Analysis of the <italic toggle="yes">Penicillium chrysogenum</italic> Velvet Protein PcVelA Identifies Methyltransferase PcLlmA as a Novel Downstream Regulator of Fungal Development

ABSTRACT Penicillium chrysogenum is the sole industrial producer of the β-lactam antibiotic penicillin, which is the most commonly used drug for treating bacterial infections. In P. chrysogenum and other filamentous fungi, secondary metabolism and morphogenesis are controlled by the highly conserved...

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Autores principales:	Kordula Becker, Sandra Ziemons, Katharina Lentz, Michael Freitag, Ulrich Kück
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	ChIP-seq PcLlmA PcVelA Penicillium chrysogenum methyltransferase protein-DNA interactions Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ea2215becb504203998149aa0778131a
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spelling	oai:doaj.org-article:ea2215becb504203998149aa0778131a2021-11-15T15:21:14ZGenome-Wide Chromatin Immunoprecipitation Sequencing Analysis of the <italic toggle="yes">Penicillium chrysogenum</italic> Velvet Protein PcVelA Identifies Methyltransferase PcLlmA as a Novel Downstream Regulator of Fungal Development10.1128/mSphere.00149-162379-5042https://doaj.org/article/ea2215becb504203998149aa0778131a2016-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00149-16https://doaj.org/toc/2379-5042ABSTRACT Penicillium chrysogenum is the sole industrial producer of the β-lactam antibiotic penicillin, which is the most commonly used drug for treating bacterial infections. In P. chrysogenum and other filamentous fungi, secondary metabolism and morphogenesis are controlled by the highly conserved multisubunit velvet complex. Here we present the first chromatin immunoprecipitation next-generation sequencing (ChIP-seq) analysis of a fungal velvet protein, providing experimental evidence that a velvet homologue in P. chrysogenum (PcVelA) acts as a direct transcriptional regulator at the DNA level in addition to functioning as a regulator at the protein level in P. chrysogenum, which was previously described. We identified many target genes that are related to processes known to be dependent on PcVelA, e.g., secondary metabolism as well as asexual and sexual development. We also identified seven PcVelA target genes that encode putative methyltransferases. Yeast two-hybrid and bimolecular fluorescence complementation analyses showed that one of the putative methyltransferases, PcLlmA, directly interacts with PcVelA. Furthermore, functional characterization of PcLlmA demonstrated that this protein is involved in the regulation of conidiosporogenesis, pellet formation, and hyphal morphology, all traits with major biotechnological relevance. IMPORTANCE Filamentous fungi are of major interest for biotechnological and pharmaceutical applications. This is due mainly to their ability to produce a wide variety of secondary metabolites, many of which are relevant as antibiotics. One of the most prominent examples is penicillin, a β-lactam antibiotic that is produced on the industrial scale by fermentation of P. chrysogenum. In recent years, the multisubunit protein complex velvet has been identified as one of the key regulators of fungal secondary metabolism and development. However, until recently, only a little has been known about how velvet mediates regulation at the molecular level. To address this issue, we performed ChIP-seq (chromatin immunoprecipitation in combination with next-generation sequencing) on and follow-up analysis of PcVelA, the core component of the velvet complex in P. chrysogenum. We demonstrate direct involvement of velvet in transcriptional control and present the putative methyltransferase PcLlmA as a new downstream factor and interaction partner of PcVelA.Kordula BeckerSandra ZiemonsKatharina LentzMichael FreitagUlrich KückAmerican Society for MicrobiologyarticleChIP-seqPcLlmAPcVelAPenicillium chrysogenummethyltransferaseprotein-DNA interactionsMicrobiologyQR1-502ENmSphere, Vol 1, Iss 4 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	ChIP-seq PcLlmA PcVelA Penicillium chrysogenum methyltransferase protein-DNA interactions Microbiology QR1-502
spellingShingle	ChIP-seq PcLlmA PcVelA Penicillium chrysogenum methyltransferase protein-DNA interactions Microbiology QR1-502 Kordula Becker Sandra Ziemons Katharina Lentz Michael Freitag Ulrich Kück Genome-Wide Chromatin Immunoprecipitation Sequencing Analysis of the <italic toggle="yes">Penicillium chrysogenum</italic> Velvet Protein PcVelA Identifies Methyltransferase PcLlmA as a Novel Downstream Regulator of Fungal Development
description	ABSTRACT Penicillium chrysogenum is the sole industrial producer of the β-lactam antibiotic penicillin, which is the most commonly used drug for treating bacterial infections. In P. chrysogenum and other filamentous fungi, secondary metabolism and morphogenesis are controlled by the highly conserved multisubunit velvet complex. Here we present the first chromatin immunoprecipitation next-generation sequencing (ChIP-seq) analysis of a fungal velvet protein, providing experimental evidence that a velvet homologue in P. chrysogenum (PcVelA) acts as a direct transcriptional regulator at the DNA level in addition to functioning as a regulator at the protein level in P. chrysogenum, which was previously described. We identified many target genes that are related to processes known to be dependent on PcVelA, e.g., secondary metabolism as well as asexual and sexual development. We also identified seven PcVelA target genes that encode putative methyltransferases. Yeast two-hybrid and bimolecular fluorescence complementation analyses showed that one of the putative methyltransferases, PcLlmA, directly interacts with PcVelA. Furthermore, functional characterization of PcLlmA demonstrated that this protein is involved in the regulation of conidiosporogenesis, pellet formation, and hyphal morphology, all traits with major biotechnological relevance. IMPORTANCE Filamentous fungi are of major interest for biotechnological and pharmaceutical applications. This is due mainly to their ability to produce a wide variety of secondary metabolites, many of which are relevant as antibiotics. One of the most prominent examples is penicillin, a β-lactam antibiotic that is produced on the industrial scale by fermentation of P. chrysogenum. In recent years, the multisubunit protein complex velvet has been identified as one of the key regulators of fungal secondary metabolism and development. However, until recently, only a little has been known about how velvet mediates regulation at the molecular level. To address this issue, we performed ChIP-seq (chromatin immunoprecipitation in combination with next-generation sequencing) on and follow-up analysis of PcVelA, the core component of the velvet complex in P. chrysogenum. We demonstrate direct involvement of velvet in transcriptional control and present the putative methyltransferase PcLlmA as a new downstream factor and interaction partner of PcVelA.
format	article
author	Kordula Becker Sandra Ziemons Katharina Lentz Michael Freitag Ulrich Kück
author_facet	Kordula Becker Sandra Ziemons Katharina Lentz Michael Freitag Ulrich Kück
author_sort	Kordula Becker
title	Genome-Wide Chromatin Immunoprecipitation Sequencing Analysis of the <italic toggle="yes">Penicillium chrysogenum</italic> Velvet Protein PcVelA Identifies Methyltransferase PcLlmA as a Novel Downstream Regulator of Fungal Development
title_short	Genome-Wide Chromatin Immunoprecipitation Sequencing Analysis of the <italic toggle="yes">Penicillium chrysogenum</italic> Velvet Protein PcVelA Identifies Methyltransferase PcLlmA as a Novel Downstream Regulator of Fungal Development
title_full	Genome-Wide Chromatin Immunoprecipitation Sequencing Analysis of the <italic toggle="yes">Penicillium chrysogenum</italic> Velvet Protein PcVelA Identifies Methyltransferase PcLlmA as a Novel Downstream Regulator of Fungal Development
title_fullStr	Genome-Wide Chromatin Immunoprecipitation Sequencing Analysis of the <italic toggle="yes">Penicillium chrysogenum</italic> Velvet Protein PcVelA Identifies Methyltransferase PcLlmA as a Novel Downstream Regulator of Fungal Development
title_full_unstemmed	Genome-Wide Chromatin Immunoprecipitation Sequencing Analysis of the <italic toggle="yes">Penicillium chrysogenum</italic> Velvet Protein PcVelA Identifies Methyltransferase PcLlmA as a Novel Downstream Regulator of Fungal Development
title_sort	genome-wide chromatin immunoprecipitation sequencing analysis of the <italic toggle="yes">penicillium chrysogenum</italic> velvet protein pcvela identifies methyltransferase pcllma as a novel downstream regulator of fungal development
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/ea2215becb504203998149aa0778131a
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Genome-Wide Chromatin Immunoprecipitation Sequencing Analysis of the <italic toggle="yes">Penicillium chrysogenum</italic> Velvet Protein PcVelA Identifies Methyltransferase PcLlmA as a Novel Downstream Regulator of Fungal Development

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