TY - CONF
T1 - Homeodomain transcription factors in the white-rot fungus Ganoderma lucidum
AU - Vandelook, Simon
AU - Van Rossum, Ulric Dominique S
AU - Stijlemans, Benoit
AU - Peeters, Eveline
PY - 2023/3/7
Y1 - 2023/3/7
N2 - Ganoderma lucidum is a white rot filamentous fungus that belongs to the Polyporales. Besides a use in traditional Asian medicine, the species has gained enormous interest for applications in the field of sustainable mycelium materials these past years. Homeodomain (Hox) transcription factors play critical roles in the regulation of development and morphogenesis processes and although this importance, not much is known about their functioning and physiological role in G. lucidum. Previously, deletion of Hox transcription factors in the model fungus Schizophyllum commune was shown to lead to an altered vegetative mycelium growth and fruiting body development. In this project, we aim to investigate putative Hox transcription factors in a G. lucidum strain used for myco-material production, using a combined in silico and experimental approach. The genome of this strain was sequenced and bioinformatic analyses enabled to predict 12 genes encoding putative Hox transcription factor-encoding genes. This included homologs of transcription factors that were previously characterized in S. commune, such as Hom2 but also Fst4. A cDNA library was prepared and a selection of 5 genes encoding homeodomain TFs were amplified indicating expression. Followed cloning of the cDNA transcripts in pET24a for heterologous expression of proteins in E. coli. Protein purification was performed using IMAC allowing further biophysical characterization. Additionally, purified proteins were used as antigen for antibody generation. Currently, a chromatin immunoprecipitation protocol is being developed for G. lucidum that enables the mapping of genome-wide binding profile of the transcription factors as a first step towards insights into the regulon and physiological functioning.
AB - Ganoderma lucidum is a white rot filamentous fungus that belongs to the Polyporales. Besides a use in traditional Asian medicine, the species has gained enormous interest for applications in the field of sustainable mycelium materials these past years. Homeodomain (Hox) transcription factors play critical roles in the regulation of development and morphogenesis processes and although this importance, not much is known about their functioning and physiological role in G. lucidum. Previously, deletion of Hox transcription factors in the model fungus Schizophyllum commune was shown to lead to an altered vegetative mycelium growth and fruiting body development. In this project, we aim to investigate putative Hox transcription factors in a G. lucidum strain used for myco-material production, using a combined in silico and experimental approach. The genome of this strain was sequenced and bioinformatic analyses enabled to predict 12 genes encoding putative Hox transcription factor-encoding genes. This included homologs of transcription factors that were previously characterized in S. commune, such as Hom2 but also Fst4. A cDNA library was prepared and a selection of 5 genes encoding homeodomain TFs were amplified indicating expression. Followed cloning of the cDNA transcripts in pET24a for heterologous expression of proteins in E. coli. Protein purification was performed using IMAC allowing further biophysical characterization. Additionally, purified proteins were used as antigen for antibody generation. Currently, a chromatin immunoprecipitation protocol is being developed for G. lucidum that enables the mapping of genome-wide binding profile of the transcription factors as a first step towards insights into the regulon and physiological functioning.
M3 - Poster
ER -