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Interactions define the dynamics of life. Electrostatic interactions provide a transient and tunable response, whereas hydrophobic interactions confer stability. Post-translational protein modifications (PTMs) such as phosphorylation, acetylation and poly(ADP-ribosyl)ation provide a regulatory switch by interfering with these interactions. Our aim is to understand how protein interactions and PTMs fine tune the dynamics of different cellular processes, in particular DNA damage response and transcription. In DNA damage response, PTMs modulate chromatin structure and promote recruitment of signalling and repair proteins. During transcription, dynamic phosphorylation of the C-terminal domain of RNA polymerase II governs timely recruitment of transcription and RNA processing factors. Revealing the function and regulation of DNA repair and transcription factors will help in understanding how their deficiency or misregulation leads to a broad range of diseases.
We apply an integrative approach to address our research questions, including biochemistry, molecular cell biology, structural biology or stem cell biology. To study protein interactions we use mass spectrometry, in vitro binding assays (isothermal titration calorimetry, fluorescence anisotropy) and FLIM-FRET (fluorescence lifetime imaging-Förster resonance energy transfer). To understand the structural basis of protein interactions, we apply X-ray crystallography, NMR and Cryo-EM through collaborative efforts. To probe protein function, we perform CRISPR/Cas9 genome editing in mammalian cells. We use live cell imaging to monitor cell cycle progression and protein recruitment to laser-induced DNA damage sites. We combine a vast array of functional genomic approaches: RNAseq and PROseq to study transcription, ChIPseq to study genomic occupancy, SLAMseq to study RNA stability. Using an arsenal of different techniques we endeavour to build a complete picture about protein function and regulation.
I obtained my BSc/MSc in Molecular Biology and Pharmacy from the University of Zagreb. I carried out my PhD thesis on DNA repair in the radiation-resistant bacterium Deinococcus radiodurans with Miroslav Radman at the University Pierre et Marie Curie in Paris. As a post-doc I joined Ivan Ahel at the Paterson Institute in Manchester to work on PARG. I started my group at Max Perutz Labs in 2012.
We identified a non-canonical PIP-box in the disordered regulatory region of poly(ADP-ribose) glycohydrolase (PARG) as the PCNA binding motif and showed based on X-ray structure analysis that the PARG PIP-box interacts with PCNA via both hydrophobic and electrostatic interactions. PARG PIP-box is critical for PARG recruitment to DNA damage sites.
Sirtuin 2 (SIRT2) is a ubiquitous mammalian NAD-dependent deacetylase. We showed that SIRT2 regulates nuclear envelope reassembly by regulating the acetylation state of ANKLE2 (Ankyrin and LEM-domain containing protein 2). SIRT2 depletion or overexpression causes nuclear envelope defects.
We advanced a laser microirradiation setup to monitor simultaneously the recruitment of two proteins to DNA damage sites. We also expanded the FLIM system with a UV laser to measure biophysical parameters of protein-protein interaction at a single cell level after DNA damage.
PHF3 regulates neuronal gene expression through the Pol II CTD reader domain SPOC.
Appel, Lisa-Marie; Franke, Vedran; Bruno, Melania; Grishkovskaya, Irina; Kasiliauskaite, Aiste; Kaufmann, Tanja; Schoeberl, Ursula E; Puchinger, Martin G; Kostrhon, Sebastian; Ebenwaldner, Carmen; Sebesta, Marek; Beltzung, Etienne; Mechtler, Karl; Lin, Gen; Vlasova, Anna; Leeb, Martin; Pavri, Rushad; Stark, Alexander; Akalin, Altuna; Stefl, Richard; Bernecky, Carrie; Djinovic-Carugo, Kristina; Slade, Dea
A novel non-canonical PIP-box mediates PARG interaction with PCNA
Tanja Kaufmann, Irina Grishkovskaya, Anton A. Polyansky, Sebastian Kostrhon, Eva Kukolj, Karin M. Olek, Sebastien Herbert, Etienne Beltzung, Karl Mechtler, Thomas Peterbauer, Josef Gotzmann, Lijuan Zhang, Markus Hartl, Bojan Zagrovic, Kareem Elsayad, Kristina Djinovic-Carugo, Dea Slade
SIRT2 regulates nuclear envelope reassembly via ANKLE2 deacetylation.
Kaufmann, Tanja; Kukolj, Eva; Brachner, Andreas; Beltzung, Etienne; Bruno, Melania; Kostrhon, Sebastian; Opravil, Susanne; Hudecz, Otto; Mechtler, Karl; Warren, Graham; Slade, Dea
PARP inhibition causes premature loss of cohesion in cancer cells.
Kukolj, Eva; Kaufmann, Tanja; Dick, Amalie E; Zeillinger, Robert; Gerlich, Daniel W; Slade, Dea
Direct measurement of protein-protein interactions by FLIM-FRET at UV laser-induced DNA damage sites in living cells.
Kaufmann, T; Herbert, S; Hackl, B; Besold, J M; Schramek, C; Gotzmann, J; Elsayad, K; Slade, D
Stand-alone P 31112-B28, Stand-alone P31546-B28
2016-2019: The Group Slade participates in the special Doctoral Program "Integrative Structural Biology" reviewed and funded by the Austrian Science Fund FWF.
The group Slade is an associated member of the special Doctoral Program "Chromosome Dynamics" reviewed and funded by the Austrian Research Fund FWF.