Universität zu Köln

Becker, Kordula, Ziemons, Sandra, Lentz, Katharina, Freitag, Michael and Kuck, Ulrich ORCID: 0000-0002-2998-8230 (2016). Genome-Wide Chromatin Immunoprecipitation Sequencing Analysis of the Penicillium chrysogenum Velvet Protein PcVelA Identifies Methyltransferase PcLlmA as a Novel Downstream Regulator of Fungal Development. mSphere, 1 (4). WASHINGTON: AMER SOC MICROBIOLOGY. ISSN 2379-5042

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Abstract

Penicillium chrysogenum is the sole industrial producer of the beta-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 beta-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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Becker, Kordula UNSPECIFIED UNSPECIFIED UNSPECIFIED Ziemons, Sandra UNSPECIFIED UNSPECIFIED UNSPECIFIED Lentz, Katharina UNSPECIFIED UNSPECIFIED UNSPECIFIED Freitag, Michael UNSPECIFIED UNSPECIFIED UNSPECIFIED Kuck, Ulrich UNSPECIFIED orcid.org/0000-0002-2998-8230 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-270182
DOI:	10.1128/mSphere.00149-16
Journal or Publication Title:	mSphere
Volume:	1
Number:	4
Date:	2016
Publisher:	AMER SOC MICROBIOLOGY
Place of Publication:	WASHINGTON
ISSN:	2379-5042
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language DEPENDENT GENE-EXPRESSION; TRANSCRIPTION FACTOR; ASPERGILLUS-NIDULANS; CONIDIOPHORE DEVELOPMENT; SECONDARY METABOLISM; TARGET GENES; I-TASSER; COMPLEX; BIOSYNTHESIS; LIGHT Multiple languages Microbiology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/27018