The Mre11-Nbs1 Interface Is Essential for Viability and Tumor Suppression

The Mre11 complex (Mre11, Rad50, and Nbs1) is integral to both DNA repair and ataxia telangiectasia mutated (ATM)-dependent DNA damage signaling. All three Mre11 complex components are essential for viability at the cellular and organismal levels. To delineate essential and non-essential Mre11 complex functions that are mediated by Nbs1, we used TALEN-based genome editing to derive Nbs1 mutant mice (Nbs1mid mice), which harbor mutations in the Mre11 interaction domain of Nbs1. Nbs1mid alleles that abolished interaction were incompatible with viability. Conversely, a 108-amino-acid Nbs1 fragment comprising the Mre11 interface was sufficient to rescue viability and ATM activation in cultured cells and support differentiation of hematopoietic cells in vivo. These data indicate that the essential role of Nbs1 is via its interaction with Mre11 and that most of the Nbs1 protein is dispensable for Mre11 complex functions and suggest that Mre11 and Rad50 directly activate ATM

The Mre11-Nbs1 interface is essential for viability and tumor suppression

The Mre11 complex (Mre11, Rad50, and Nbs1) is integral to both DNA repair and ataxia telangiectasia mutated (ATM)-dependent DNA damage signaling. All three Mre11 complex components are essential for viability at the cellular and organismal levels. To delineate essential and non-essential Mre11 complex functions that are mediated by Nbs1, we used TALEN-based genome editing to derive Nbs1 mutant mice (Nbs1mid mice), which harbor mutations in the Mre11 interaction domain of Nbs1. Nbs1mid alleles that abolished interaction were incompatible with viability. Conversely, a 108-amino-acid Nbs1 fragment comprising the Mre11 interface was sufficient to rescue viability and ATM activation in cultured cells and support differentiation of hematopoietic cells in vivo. These data indicate that the essential role of Nbs1 is via its interaction with Mre11 and that most of the Nbs1 protein is dispensable for Mre11 complex functions and suggest that Mre11 and Rad50 directly activate ATM

Architectural plasticity of human BRCA2-RAD51 complexes in DNA break repair

The tumor suppressor BRCA2 is a large multifunctional protein mutated in 50-60% of familial breast cancers. BRCA2 interacts with many partners and includes multiple regions with potentially disordered structure. In homology directed DNA repair BRCA2 delivers RAD51 to DNA resulting in removal of RPA and assembly of a RAD51 nucleoprotein filame

Structural dynamics of a nanomachine: investigating structure-to-function properties of large, flexible, DNA-associating proteins

Proteins, the working tools of each and every cell, are able to carry out and catalyse all processes essential for cellular metabolism and survival. Every protein is equipped with a set of specific properties, e.g. catalytic or structural features, that enable them to fulfil specific tasks necessary for the proper functioning of the cell. Moreover, proteins and peptides can assemble in intricate, multicomponent arrays, that further increases the efficiency and specificity of performed tasks, thus functioning as biological nanomachines. As in every machine, or tool, structure and structural dynamics are inseparable with protein’ function. With the recent developments in techniques utilized to study protein structure, advances in understanding protein function had been remarkable, however, big, flexible proteins prove difficult to study and very often remain outside the scope of conventional biochemical investigations. The aim of this thesis was to employ Scanning Force Microscopy (SFM) to investigate structure-to- function relations of proteins which size and structural dynamics are limiting factors in most biochemical approaches

Enhanced Chromatin Dynamics by FACT Promotes Transcriptional Restart after UV-Induced DNA Damage

Chromatin remodeling is tightly linked to all DNA-transacting activities. To study chromatin remodeling during DNA repair, we established quantitative fluorescence imaging methods to measure the exchange of histones in chromatin in living cells. We show that particularly H2A and H2B are evicted and replaced at an accelerated pace at sites of UV-induced DNA damage. This accelerated exchange of H2A/H2B is facilitated by SPT16, one of the two subunits of the histone chaperone FACT (facilitates chromatin transcription) but largely independent of its partner SSRP1. Interestingly, SPT16 is targeted to sites of UV light-induced DNA damage-arrested transcription and is required for efficient restart of RNA synthesis upon damage removal. Together, our data uncover an important role for chromatin dynamics at the crossroads of transcription and the UV-induced DNA damage response

Toward a pulsed antihydrogen beam for WEP tests in AEgIS

The AEg̅IS collaboration at CERN’s AD produces antihydrogen atoms in the form of a pulsed, isotropic source with a precisely defined formation time. AEg̅IS has recently undergone major upgrades to fully benefit from the increased number of colder antiprotons provided by the new ELENA decelerator and to move toward forming a horizontal beam to directly investigate the influence of gravity on the H̅ atoms, thereby probing the Weak Equivalence Principle for antimatter. This contribution gives an overview of these upgrades as well as subsequent results from the first beam times with ELENA