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

Positronium laser cooling via the 13S1^3S-23P2^3P transition with a broadband laser pulse

We report on laser cooling of a large fraction of positronium (Ps) in free-flight by strongly saturating the $1^3S$-$2^3P$ transition with a broadband, long-pulsed 243 nm alexandrite laser. The ground state Ps cloud is produced in a magnetic and electric field-free environment. We observe two different laser-induced effects. The first effect is an increase in the number of atoms in the ground state after the time Ps has spent in the long-lived $3^3P$ states. The second effect is the one-dimensional Doppler cooling of Ps, reducing the cloud's temperature from 380(20) K to 170(20) K. We demonstrate a 58(9) % increase in the coldest fraction of the Ps ensemble.Comment: 6 pages, 5 figure

Positronium Laser Cooling via the 1 3 S − 2 3 P Transition with a Broadband Laser Pulse

We report on laser cooling of a large fraction of positronium (Ps) in free flight by strongly saturating the 1^{3}S-2^{3}P transition with a broadband, long-pulsed 243 nm alexandrite laser. The ground state Ps cloud is produced in a magnetic and electric field-free environment. We observe two different laser-induced effects. The first effect is an increase in the number of atoms in the ground state after the time Ps has spent in the long-lived 2^{3}P states. The second effect is one-dimensional Doppler cooling of Ps, reducing the cloud's temperature from 380(20) to 170(20) K. We demonstrate a 58(9)% increase in the fraction of Ps atoms with v_{1D}<3.7×10^{4}  ms^{-1}

CIRCUS: an autonomous control system for antimatter, atomic and quantum physics experiments

AbstractA powerful and robust control system is a crucial, often neglected, pillar of any modern, complex physics experiment that requires the management of a multitude of different devices and their precise time synchronisation. The AEḡIS collaboration presents CIRCUS, a novel, autonomous control system optimised for time-critical experiments such as those at CERN’s Antiproton Decelerator and, more broadly, in atomic and quantum physics research. Its setup is based on Sinara/ARTIQ and TALOS, integrating the ALPACA analysis pipeline, the last two developed entirely in AEḡIS. It is suitable for strict synchronicity requirements and repeatable, automated operation of experiments, culminating in autonomous parameter optimisation via feedback from real-time data analysis. CIRCUS has been successfully deployed and tested in AEḡIS; being experiment-agnostic and released open-source, other experiments can leverage its capabilities.</jats:p

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

CIRCUS: an autonomous control system for antimatter, atomic and quantum physics experiments

A powerful and robust control system is a crucial, often neglected, pillar of any modern, complex physics experiment that requires the management of a multitude of different devices and their precise time synchronisation. The AEḡIS collaboration presents CIRCUS, a novel, autonomous control system optimised for time-critical experiments such as those at CERN’s Antiproton Decelerator and, more broadly, in atomic and quantum physics research. Its setup is based on Sinara/ARTIQ and TALOS, integrating the ALPACA analysis pipeline, the last two developed entirely in AEḡIS. It is suitable for strict synchronicity requirements and repeatable, automated operation of experiments, culminating in autonomous parameter optimisation via feedback from real-time data analysis. CIRCUS has been successfully deployed and tested in AEḡIS; being experiment-agnostic and released open-source, other experiments can leverage its capabilities

Pulsed Production of Antihydrogen in AEgIS

Low-temperature antihydrogen atoms are an effective tool to probe the validity of the fundamental laws of Physics, for example the Weak Equivalence Principle (WEP) for antimatter, and -generally speaking- it is obvious that colder atoms will increase the level of precision. After the first production of cold antihydrogen in 2002 [1], experimental efforts have substantially progressed, with really competitive results already reached by adapting to cold antiatoms some well-known techniques pre- viously developed for ordinary atoms. Unfortunately, the number of antihydrogen atoms that can be produced in dedicated experiments is many orders of magnitude smaller than of hydrogen atoms, so the development of novel techniques to enhance the production of antihydrogen with well defined (and possibly controlled) conditions is essential to improve the sensitivity. We present here some experimental results achieved by the AEgIS Collaboration, based at the CERN AD (Antiproton Decelerator) on the production of antihydrogen in a pulsed mode where the production time of 90% of atoms is known with an uncertainty of ~ 250 ns [2]. The pulsed antihydrogen source is generated by the charge-exchange reaction between Rydberg positronium (Ps*) and an antiproton (p¯): p¯ + Ps* → H¯* + e−, where Ps* is produced via the implantation of a pulsed positron beam into a mesoporous silica target, and excited by two consecutive laser pulses, and antiprotons are trapped, cooled and manipulated in Penning-Malmberg traps. The pulsed production (which is a major milestone for AEgIS) makes it possible to select the antihydrogen axial temperature and opens the door for the tuning of the antihydrogen Rydberg states, their de-excitation by pulsed lasers and the manipulation through electric field gradients. In this paper, we present the results achieved by AEgIS in 2018, just before the Long Shutdown 2 (LS2), as well as some of the ongoing improvements to the system, aimed at exploiting the lower energy antiproton beam from ELENA [3].</jats:p

Positronium laser cooling via the 13S1^3S-23P2^3P transition with a broadband laser pulse

We report on laser cooling of a large fraction of positronium (Ps) in free-flight by strongly saturating the $1^3S$-$2^3P$ transition with a broadband, long-pulsed 243 nm alexandrite laser. The ground state Ps cloud is produced in a magnetic and electric field-free environment. We observe two different laser-induced effects. The first effect is an increase in the number of atoms in the ground state after the time Ps has spent in the long-lived $3^3P$ states. The second effect is the one-dimensional Doppler cooling of Ps, reducing the cloud's temperature from 380(20) K to 170(20) K. We demonstrate a 58(9) % increase in the coldest fraction of the Ps ensemble