Slx5/Slx8-dependent ubiquitin hotspots on chromatin contribute to stress tolerance

Chromatin is a highly regulated environment, and protein association with chromatin is often controlled by post-translational modifications and the corresponding enzymatic machinery. Specifically, SUMO-targeted ubiquitin ligases (STUbLs) have emerged as key players in nuclear quality control, genome maintenance, and transcription. However, how STUbLs select specific substrates among myriads of SUMOylated proteins on chromatin remains unclear. Here, we reveal a remarkable co-localization of the budding yeast STUbL Slx5/Slx8 and ubiquitin at seven genomic loci that we term "ubiquitin hotspots". Ubiquitylation at these sites depends on Slx5/Slx8 and protein turnover on the Cdc48 segregase. We identify the transcription factor-like Ymr111c/Euc1 to associate with these sites and to be a critical determinant of ubiquitylation. Euc1 specifically targets Slx5/Slx8 to ubiquitin hotspots via bipartite binding of Slx5 that involves the Slx5 SUMO-interacting motifs and an additional, novel substrate recognition domain. Interestingly, the Euc1-ubiquitin hotspot pathway acts redundantly with chromatin modifiers of the H2A.Z and Rpd3L pathways in specific stress responses. Thus, our data suggest that STUbL-dependent ubiquitin hotspots shape chromatin during stress adaptation

LRRC8/VRAC anion channels enhance β-cell glucose sensing and insulin secretion

Glucose homeostasis depends critically on insulin that is secreted by pancreatic β-cells. Serum glucose, which is directly sensed by β-cells, stimulates depolarization- and Ca(2+)-dependent exocytosis of insulin granules. Here we show that pancreatic islets prominently express LRRC8A and LRRC8D, subunits of volume-regulated VRAC anion channels. Hypotonicity- or glucose-induced β-cell swelling elicits canonical LRRC8A-dependent VRAC currents that depolarize β-cells to an extent that causes electrical excitation. Glucose-induced excitation and Ca(2+) responses are delayed in onset, but not abolished, in β-cells lacking the essential VRAC subunit LRRC8A. Whereas Lrrc8a disruption does not affect tolbutamide- or high-K(+)-induced insulin secretion from pancreatic islets, it reduces first-phase glucose-induced insulin secretion. Mice lacking VRAC in β-cells have normal resting serum glucose levels but impaired glucose tolerance. We propose that opening of LRRC8/VRAC channels increases glucose sensitivity and insulin secretion of β-cells synergistically with K(ATP) closure. Neurotransmitter-permeable LRRC8D-containing VRACs might have additional roles in autocrine/paracrine signaling within islets

Versatile compact atomic source for high resolution dual atom interferometry

We present a compact $^{87}$Rb atomic source for high precision dual atom interferometers. The source is based on a double-stage magneto-optical trap (MOT) design, consisting of a 2-dimensional (2D)-MOT for efficient loading of a 3D-MOT. The accumulated atoms are precisely launched in a horizontal moving molasses. Our setup generates a high atomic flux ($>10^{10}$ atoms/s) with precise and flexibly tunable atomic trajectories as required for high resolution Sagnac atom interferometry. We characterize the performance of the source with respect to the relevant parameters of the launched atoms, i.e. temperature, absolute velocity and pointing, by utilizing time-of-flight techniques and velocity selective Raman transitions.Comment: uses revtex4, 9 pages, 12 figures, submitted to Phys. Rev.

Inactivation and anion selectivity of volume-regulated VRAC channels depend on carboxy-terminal residues of the first extracellular loop

Canonical volume-regulated anion channels (VRACs) are crucial for cell volume regulation and have many other important roles including tumor drug resistance and release of neurotransmitters. Although VRAC-mediated swelling-activated chloride currents (ICl,vol) have been studied for decades, exploration of the structure-function relationship of VRAC has become possible only after the recent discovery that VRACs are formed by differently composed heteromers of LRRC8 proteins. Inactivation of ICl,vol at positive potentials, a typical hallmark of VRACs, strongly varies between native cell types. Exploiting the large differences in inactivation between different LRRC8 heteromers, we now used chimeras assembled from isoforms LRRC8C and LRRC8E to uncover a highly conserved extracellular region preceding the second LRRC8 transmembrane domain as a major determinant of ICl,vol inactivation. Point mutations identified two amino-acids (Lys98 and Asp100 in LRRC8A and equivalent residues in LRRC8C and -E) which upon charge reversal strongly altered the kinetics and voltage-dependence of inactivation. Importantly, charge reversal at the first position also reduced the iodide>chloride permeability of ICl,vol. This change in selectivity was stronger when both the obligatory LRRC8A subunit and the other co-expressed isoform (LRR8C or -E) carried such mutations. Hence the carboxy-terminal part of the first extracellular loop not only determines VRAC inactivation, but might also participate in forming its outer pore. Inactivation of VRACs may involve a closure of the extracellular mouth of the permeation pathway

Gauss sum factorization with cold atoms

We report the first implementation of a Gauss sum factorization algorithm by an internal state Ramsey interferometer using cold atoms. A sequence of appropriately designed light pulses interacts with an ensemble of cold rubidium atoms. The final population in the involved atomic levels determines a Gauss sum. With this technique we factor the number N=263193.Comment: 4 pages, 5 figure

Assessing the risk of climate change for buildings: A comparison between multi-year and probabilistic reference year simulations

Copyright © 2011 Elsevier. NOTICE: this is the author’s version of a work that was accepted for publication in Building and Environment . Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Building and Environment Vol. 46 (2011), DOI: 10.1016/j.buildenv.2010.12.018Given a changing climate, there is a need to provide data for future years so that practicing engineers can investigate the impact of climate change on particular designs and examine any risk the client might be exposed to. In addition, such files are of use to building scientists in developing generic solutions to problems such as elevated internal temperatures and poor thermal comfort. With the release of the UK Climate Projections (UKCP09) [1], and the publication of a methodology for the creation of probabilistic future reference years using the UKCP09 weather generator [2], it is possible to model future building performance. However, the collapse of the distribution of possibilities inherent in the UKCP09 method into a single reference year or a small number of reference years, potentially means the loss of most of the information about the potential range of the response of the building and of the risk occupants might be subject to. In this paper we model for the first time the internal conditions and energy use of a building with all 3000 example years produced by the UKCP09 weather generator in an attempt to study the full range of response and risk. The resultant histograms and cumulative distribution functions are then used to examine whether single reference years can be used to answer questions about response and risk under a changing climate, or whether a more probabilistic approach is unavoidable

Differential atom interferometry beyond the standard quantum limit

We analyze methods to go beyond the standard quantum limit for a class of atomic interferometers, where the quantity of interest is the difference of phase shifts obtained by two independent atomic ensembles. An example is given by an atomic Sagnac interferometer, where for two ensembles propagating in opposite directions in the interferometer this phase difference encodes the angular velocity of the experimental setup. We discuss methods of squeezing separately or jointly observables of the two atomic ensembles, and compare in detail advantages and drawbacks of such schemes. In particular we show that the method of joint squeezing may improve the variance by up to a factor of 2. We take into account fluctuations of the number of atoms in both the preparation and the measurement stage, and obtain bounds on the difference of the numbers of atoms in the two ensembles, as well as on the detection efficiency, which have to be fulfilled in order to surpass the standard quantum limit. Under realistic conditions, the performance of both schemes can be improved significantly by reading out the phase difference via a quantum non-demolition (QND) measurement. Finally, we discuss a scheme using macroscopically entangled ensembles.Comment: 10 pages, 5 figures; eq. (3) corrected and other minor change

Identification of TMEM206 proteins as pore of PAORAC/ASOR acid-sensitive chloride channels

Acid-sensing ion channels have important functions in physiology and pathology, but the molecular composition of acid-activated chloride channels had remained unclear. We now used a genome-wide siRNA screen to molecularly identify the widely expressed acid-sensitive outwardly-rectifying anion channel PAORAC/ASOR. ASOR is formed by TMEM206 proteins which display two transmembrane domains (TMs) and are expressed at the plasma membrane. Ion permeation-changing mutations along the length of TM2 and at the end of TM1 suggest that these segments line ASOR's pore. While not belonging to a gene family, TMEM206 has orthologs in probably all vertebrates. Currents from evolutionarily distant orthologs share activation by protons, a feature essential for ASOR's role in acid-induced cell death. TMEM206 defines a novel class of ion channels. Its identification will help to understand its physiological roles and the diverse ways by which anion-selective pores can be formed