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Research project (§ 26 & § 27)
Duration : 2018-10-01 - 2019-10-31

Multiresistant microorgansims and tumors represent a continuous challenge to modern medicine. To encounter this situation, new pharmaceuticals have to be discovered and developed. A process that requires new, flexible strategies. Many pharmaceuticals originate from the kingdom of fungi. Bioinformatic analysis suggest that many more are still being hidden, even within exhaustively studied species. With this project, we want to activate these “silent” gene clusters with combination of the RNA-guided state-of-the-art tool CRISPR/CAS and an “artificial” activator, composed of viral and human transactivation domains. The system has already been established in other organisms and is therefore the tool of choice for our project. To our knowledge, there is no literature about the activation of “silent” gene clusters in filamentous fungi to this date. Should this project succeed, it would open completely new opportunities for the research of bioactive substances. Furthermor it would render the BOKU as an attractive partner for upcoming collaborations with pharmaceutical companies, who are interested in the discovery, development and production of new bioactive compounds, suitable for pharmaceutical industry.
Research project (§ 26 & § 27)
Duration : 2018-08-01 - 2021-07-31

The life of a plant is a permanent response to environmental stimuli. Plants monitor and constantly integrate the environmental fluctuations in order to adjust their growth and development. Plant hormones are central to these adaptive growth responses. Auxin is a major plant hormone that mediates a plethora of developmental responses in a concentration-dependent manner. Importantly, auxin mediates high temperature (HT)-related responses in root, shoot, and during reproductive development. PHYTOCHROME INTERACTING FACTOR 4 (PIF4) mediates auxin-dependent hypocotyl or petioles elongation to HT. In contrast, root response to HT is independent of PIF4 and hence the impact of auxin to HT-induced root growth is controversial. Similarly, the role of auxin in regulating female floral organ development under HT has not yet been thoroughly investigated. Facing the consequences of global warming, plant growth response to HT is a timely, fundamental research topic with additional potential for applied research fields. Thus, in this research, I propose to investigate the auxin-based mechanisms regulating plant organ response to HT. I present here preliminary results and propose further experiments, covering two research directions: -The first part (A) is the continuation of my submitted (preprinted) manuscript that revealed the role of PILS6 in auxin-dependent root growth under HT. I propose in this part that the transcription factor SPATULA (SPT) is involved in this and aim to investigate how HT impacts on this SPT-PILS6 module to regulate nuclear auxin signalling during root growth. This research will clarify some controversy in the literature and will enable me to dissect whether the hypocotyl (PIF4-dependent) and roots (SPT-dependent) underlie distinct auxin-dependent mechanisms. -The second part (B) will assess how PILS-dependent intracellular auxin signalling affects female flower organ development and productivity under HT conditions. This research will advance the mechanistic understanding of female floral organ development, the pollen-pistil interaction, and the potential role of auxin to regulate these developmental aspects under HT condition. My research direction will fill eminent knowledge gaps and solve some controversy in our understanding of auxin-mediated thermo-responses. This project will contribute to several research fields, connecting temperature sensing, intracellular auxin transport and plant adaptive growth under HT. Overall, my results will further our understanding of how plants respond to HT.
Research project (§ 26 & § 27)
Duration : 2018-11-01 - 2022-10-31

O-Glycosylation of nuclear and cytosolic proteins is a very common post-translational modification (PTM), contributing to the complexity in the function and regulation of proteins. In contrast to other types of glycosylation, a single sugar residue - either N-acetylglucosamine (GlcNAc) or fucose - is attached to serine or threonine residues of a high number of very diverse proteins. O-GlcNAc modification is well characterized in animals, where it is essential for development and decisively involved in a range of signaling events, often affecting protein interactions and competing with other O-linked PTMs. While animals carry only one O-GlcNAc-transferase (OGT), plants have two structurally related O-glycosyltransferases, competing for the same targets: the O-GlcNAc transferase SECRET AGENT (SEC) and the protein O-fucosyltransferase SPINDLY (SPY) show high sequence homology, but their activity can have opposite effects on their target proteins. This mechanism of counteracting O-glycosyltransferases seems to be specific for plants. Its importance for plant development is apparent from genetic analysis, as a double knockout of the two enzymes is embryonic lethal. However, clearly more research is necessary in order to understand the molecular mechanisms underlying O-glycosylation in plants. We are currently working on this topic using a combination of different approaches, including identification of targets of SPY and SEC, as well as interacting proteins. Among the proteins identified in these experiments was a group of jacalin-like lectins, that potentially interact with O-glycosylated proteins and might represent an additional regulatory layer by specifically recognizing O-GlcNAc and/or O-fucose-modified proteins, thereby mediating the molecular effects of O-glycosylation. The current proposal is therefore focused on characterization of these jacalin-like lectins and their interaction with glycoproteins. In addition, I am proposing an unbiased approach to identify new plant lectins specific for this type of O-glycosylation. The suggested experiments include testing the interaction of identified lectins with O-glycosylated proteins and determining their specificity for certain carbohydrates on glycan arrays. In parallel, the biological role of these lectins will be determined by phenotypic analysis of mutants as well as standard molecular biology techniques. This approach might elucidate the role of previously uncharacterized lectins, potentially involved in fine-tuning the effects of O-glycosylation, for instance in response to environmental stimuli. On the other hand, the identification and characterization of new lectins might be the basis for the development of better tools to analyze O-glycosylation specifically, in plants as well as other organisms.

Supervised Theses and Dissertations