Genetics of Eukaryotic Microorganisms
Molecular Biology of Fungi 2022
In September 2022 the 14th Symposium 'Molecular Biology of Fungi' took place at the Technical University Kaiserslautern. In this conference, organized by the VAAM special group 'Biology and Biotechnology of Fungi' and the GenAG 'Fungal Genetics' of the Society of Genetics, multi facets of current research regarding filamentous fungi were presented. Highlights for our group were the talk of Lucas Hollstein about his establishment of a protein-proximity labeling approach and the poster prize awarded to Anina Rudolph for her work about 'Spore types-specific infection of different maize tissues by Colletotrichum graminicola.
Complete article (BIOspektrum).
Recent publication in Scientific Reports: Establishment of in vivo proximity labeling with biotin using TurboID in the flamentous fungus Sordaria macrospora
Proximity-dependent biotin identifcation (BioID) has emerged as a powerful methodology to identify proteins co-localizing with a given bait protein in vivo. The approach has been established in animal cells, plants and yeast but not yet in flamentous fungi. BioID relies on promiscuous biotin ligases fused to bait proteins to covalently label neighboring proteins with biotin. Biotinylated proteins are specifcally enriched through biotin affnity capture from denatured cell lysates and subsequently identifed and quantifed with liquid chromatography-mass spectrometry (LC–MS). In contrast to many other affnity capture approaches for studying protein–protein interactions, BioID does not rely on physical protein–protein binding within native cell lysates. This feature allows the identifcation of protein proximities of weak or transient and dynamic nature. Here, we demonstrate the application of BioID for the fungal model organism Sordaria macrospora (Sm) using the example of the STRIPAK complex interactor 1 (SCI1) of the well-characterized striatin-interacting phosphatase and kinase (SmSTRIPAK) complex as proof of concept. For the establishment of BioID in S. macrospora, a codon-optimized TurboID biotin ligase was fused to SCI1. Biotin capture of the known SmSTRIPAK components PRO11, SmMOB3, PRO22 and SmPP2Ac1 demonstrates the successful BioID application in S. macrospora. BioID proximity labeling approaches will provide a powerful proteomics tool for fungal biologists.
more…Research - Sordaria macrospora and Colletotrichum graminicola
Filamentous fungi are great model organisms to study fundamental processes in eukaryotes as well as plant pathogen interactions. Our group focuses on both aspects of fungal life, investigating autophagy and multicellular development in Sordaria macrospora and Colletotrichum graminicola interaction with its host zea mays.
Sordaria macrospora
Fruiting-body development in filamentous ascomycetes is a complex cellular differentiation process that requires special environmental conditions and is controlled by many developmentally regulated genes. We are interested in the genes regulating this development process. We use the homothallic (self-fertile) ascomycete Sordaria macrospora as a model organism. Since S. macrospora is able to complete the sexual cycle without a mating partner, recessive mutations affecting fruiting body development are directly visible. Numerous mutants which are blocked at various stages of fruiting-body development have been generated and molecular genetic procedures have been applied to isolate genes involved in fruiting-body. more...
Colletotrichum graminicola
Cellular communication processes within a fungal colony or in host-pathogen interactions are crucial to allow adequate development or host infection. We use the hemibiotrophic maize pathogen Colletotrichum graminicola to study both communication types with special emphasis on germling fusion, sexual development, chemotropic growth to host plant signals, and plant infection. Seeking for the identification of signals, receptors and downstream pathways, we apply genetic, biochemical, bioanalytical, and microscopy techniques. Based on these findings, we also evaluate communication factors for their probable use in biotechnological applications. more...