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Kordula Becker, Sandra Ziemons, Katharina Lentz, Michael Freitag, Ulrich Kück

• \(\textit {Penicillium chrysogenum}\) is the sole industrial producer of the \(\beta\)-lactam antibiotic penicillin, which is the most commonly used drug for treating bacterial infections. In \(\textit {P. chrysogenum}\) 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 \(\textit {P. chrysogenum}\) (PcVelA) acts as a direct transcriptional regulator at the DNA level in addition to functioning as a regulator at the protein level in \(\textit {P. chrysogenum}\), 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.