Characterization of the major Woronin body protein HexA of the human pathogenic mold Aspergillus fumigatus.

In filamentous fungi, the septal pore controls the exchange between neighbouring hyphal compartments. Woronin bodies are fungal-specific organelles that plug the pore in case of physical damage. The Hex protein is their major and essential component. Hex proteins of different size are predicted in the data base for pathogenic and non-pathogenic Aspergillus species. However, using specific monoclonal antibodies, we identified 2 dominant HexA protein species of 20 and 25kDa in A. fumigatus, A. terreus, A. nidulans, and A. oryzae. HexA and Woronin bodies were found in A. fumigatus hyphae, but also in resting conidia. Using monoclonal antibodies, a GFP-HexA fusion protein, and an RFP protein fused to the putative peroxisomal targeting sequence of HexA, we analyzed the spatial localization and dynamics of Woronin bodies in A. fumigatus as well as their formation from peroxisomes. In intact hyphae, some Woronin bodies were found in close proximity to the septal pore, while the majority was distributed in the cytoplasm. Septum-associated Woronin bodies show a minimal lateral movement, while the cytosolic Woronin bodies are highly dynamic. The distribution of Woronin bodies and their co-localization pattern with peroxisomes revealed no evidence that Woronin bodies arise predominantly at the apical tip of A. fumigatus hyphae. We found that Woronin bodies are able to plug septal pores of A. fumigatus in case of damage. Woronin bodies therefore contribute to the stress resistance and potentially also to the virulence of A. fumigatus, which renders them a potential target for future anti-fungal strategies.