Structural Dynamics of the YidC:Ribosome Complex during Membrane Protein Biogenesis

Members of the YidC/Oxa1/Alb3 family universally facilitate membrane protein biogenesis, via mechanisms that have thus far remained unclear. Here, we investigated two crucial functional aspects: the interaction of YidC with ribosome: nascent chain complexes (RNCs) and the structural dynamics of RNC-bound YidC in nanodiscs. We observed that a fully exposed nascent transmembrane domain (TMD) is required for high-affinity YidC: RNC interactions, while weaker binding may already occur at earlier stages of translation. YidC efficiently catalyzed the membrane insertion of nascent TMDs in both fluid and gel phase membranes. Cryo-electron microscopy and fluorescence analysis revealed a conformational change in YidC upon nascent chain insertion: the essential TMDs 2 and 3 of YidC were tilted, while the amphipathic helix EH1 relocated into the hydrophobic core of the membrane. We suggest that EH1 serves as a mechanical lever, facilitating a coordinated movement of YidC TMDs to trigger the release of nascent chains into the membrane

The Cluster Mass Function from Early SDSS Data: Cosmological Implications

The mass function of clusters of galaxies is determined from 400 deg^2 of early commissioning imaging data of the Sloan Digital Sky Survey; ~300 clusters in the redshift range z = 0.1 - 0.2 are used. Clusters are selected using two independent selection methods: a Matched Filter and a red-sequence color magnitude technique. The two methods yield consistent results. The cluster mass function is compared with large-scale cosmological simulations. We find a best-fit cluster normalization relation of sigma_8*omega_m^0.6 = 0.33 +- 0.03 (for 0.1 ~< omega_m ~< 0.4), or equivalently sigma_8 = (0.16/omega_m)^0.6. The amplitude of this relation is significantly lower than the previous canonical value, implying that either omega_m is lower than previously expected (omega_m = 0.16 if sigma_8 = 1) or sigma_8 is lower than expected (sigma_8 = 0.7 if omega_m = 0.3). The best-fit mass function parameters are omega_m = 0.19 (+0.08,-0.07) and sigma_8 = 0.9 (+0.3,-0.2). High values of omega_m (>= 0.4) and low sigma_8 (=~ 2 sigma.Comment: AASTeX, 25 pages, including 7 figures, accepted for publication in ApJ, vol.585, March 200

Reply to: On the statistical foundation of a recent single molecule FRET benchmark

In their ‘Matters Arising’ manuscript, Saurabh et al. discuss two issues related to single-molecule Förster resonance energy transfer (smFRET) experiments: the use of the Gaussian noise approximation and spectral crosstalk. Their arguments are based on simulations obtained with parameters that differ significantly from the typical conditions measured experimentally, and, thus, from the regime included in the original study (Götz et al.1). In addition, they make claims about our multi-lab blind study that we would like to rectify. In Table 1, we provide a list of specific statements made by Saurabh et al. with our respective explanations.In our reply, we will discuss three points, summarized here and detailed below. 1.smFRET trajectories from typical surface-tethered experiments are well described by Gaussian noise models (mean photon counts are >50 per data point). Non-Gaussian Poisson noise only becomes relevant for smFRET data with extremely low photon counts, which is generally avoided by increasing the laser power and/or integration time of the experiment. 2. Spectral crosstalk correction is relevant for determining correct FRET efficiencies and FRET-derived distances, but it does not impact the kinetic rate derivation, which is the focus of Götz et al. 3. The study of Götz et al. compares the strengths and weaknesses of currently available kinetic tools to draw lessons for further development. It does not “favor” any approaches or “lead to bias” etc. as incorrectly stated by Saurabh et al. Saurabh et al. are welcome to conduct dedicated studies on the specific features they propose to extend the work of Götz et al

Cosmological parameters from SDSS and WMAP

We measure cosmological parameters using the three-dimensional power spectrum P(k) from over 200,000 galaxies in the Sloan Digital Sky Survey (SDSS) in combination with WMAP and other data. Our results are consistent with a ``vanilla'' flat adiabatic Lambda-CDM model without tilt (n=1), running tilt, tensor modes or massive neutrinos. Adding SDSS information more than halves the WMAP-only error bars on some parameters, tightening 1 sigma constraints on the Hubble parameter from h~0.74+0.18-0.07 to h~0.70+0.04-0.03, on the matter density from Omega_m~0.25+/-0.10 to Omega_m~0.30+/-0.04 (1 sigma) and on neutrino masses from <11 eV to <0.6 eV (95%). SDSS helps even more when dropping prior assumptions about curvature, neutrinos, tensor modes and the equation of state. Our results are in substantial agreement with the joint analysis of WMAP and the 2dF Galaxy Redshift Survey, which is an impressive consistency check with independent redshift survey data and analysis techniques. In this paper, we place particular emphasis on clarifying the physical origin of the constraints, i.e., what we do and do not know when using different data sets and prior assumptions. For instance, dropping the assumption that space is perfectly flat, the WMAP-only constraint on the measured age of the Universe tightens from t0~16.3+2.3-1.8 Gyr to t0~14.1+1.0-0.9 Gyr by adding SDSS and SN Ia data. Including tensors, running tilt, neutrino mass and equation of state in the list of free parameters, many constraints are still quite weak, but future cosmological measurements from SDSS and other sources should allow these to be substantially tightened.Comment: Minor revisions to match accepted PRD version. SDSS data and ppt figures available at http://www.hep.upenn.edu/~max/sdsspars.htm

The First Post-Kepler Brightness Dips of KIC 8462852

We present a photometric detection of the first brightness dips of the unique variable star KIC 8462852 since the end of the Kepler space mission in 2013 May. Our regular photometric surveillance started in October 2015, and a sequence of dipping began in 2017 May continuing on through the end of 2017, when the star was no longer visible from Earth. We distinguish four main 1-2.5% dips, named "Elsie," "Celeste," "Skara Brae," and "Angkor", which persist on timescales from several days to weeks. Our main results so far are: (i) there are no apparent changes of the stellar spectrum or polarization during the dips; (ii) the multiband photometry of the dips shows differential reddening favoring non-grey extinction. Therefore, our data are inconsistent with dip models that invoke optically thick material, but rather they are in-line with predictions for an occulter consisting primarily of ordinary dust, where much of the material must be optically thin with a size scale <<1um, and may also be consistent with models invoking variations intrinsic to the stellar photosphere. Notably, our data do not place constraints on the color of the longer-term "secular" dimming, which may be caused by independent processes, or probe different regimes of a single process

Monitoring integrity and localization of modified single-stranded RNA oligonucleotides using ultrasensitive fluorescence methods

Short single-stranded oligonucleotides represent a class of promising therapeutics with diverse application areas. Antisense oligonucleotides, for example, can interfere with various processes involved in mRNA processing through complementary base pairing. Also RNA interference can be regulated by antagomirs, single-stranded siRNA and single-stranded microRNA mimics. The increased susceptibility to nucleolytic degradation of unpaired RNAs can be counteracted by chemical modification of the sugar phosphate backbone. In order to understand the dynamics of such single-stranded RNAs, we investigated their fate after exposure to cellular environment by several fluorescence spectroscopy techniques. First, we elucidated the degradation of four differently modified, dual-dye labeled short RNA oligonucleotides in HeLa cell extracts by fluorescence correlation spectroscopy, fluorescence cross-correlation spectroscopy and Forster resonance energy transfer. We observed that the integrity of the oligonucleotide sequence correlates with the extent of chemical modifications. Furthermore, the data showed that nucleolytic degradation can only be distinguished from unspecific effects like aggregation, association with cellular proteins, or intramolecular dynamics when considering multiple measurement and analysis approaches. We also investigated the localization and integrity of the four modified oligonucleotides in cultured HeLa cells using fluorescence lifetime imaging microscopy. No intracellular accumulation could be observed for unmodified oligonucleotides, while completely stabilized oligonucleotides showed strong accumulation within HeLa cells with no changes in fluorescence lifetime over 24 h. The integrity and accumulation of partly modified oligonucleotides was in accordance with their extent of modification. In highly fluorescent cells, the oligonucleotides were transported to the nucleus. The lifetime of the RNA in the cells could be explained by a balance between release of the oligonucleotides from endosomes, degradation by RNases and subsequent depletion from the cells

Improving Access to Mental Health Care in an Orthodox Jewish Community: A Critical Reflection Upon the Accommodation of Otherness

The English National Health Service (NHS) has significantly extended the supply of evidence based psychological interventions in primary care for people experiencing common mental health problems. Yet despite the extra resources, the accessibility of services for ‘under-served’ ethnic and religious minority groups, is considerably short of the levels of access that may be necessary to offset the health inequalities created by their different exposure to services, resulting in negative health outcomes. This paper offers a critical reflection upon an initiative that sought to improve access to an NHS funded primary care mental health service to one ‘under-served’ population, an Orthodox Jewish community in the North West of England

1982-09-30 Unfinished Business: The Dry Reaction To The Wet Vote

On this episode of Unfinished Business an in depth look at the dry\u27s reaction to Morehead going wet (allowing alcohol sales) with Tim Roberts and Dave Farmer interviewing various members of the dry forces, including Roy White, Steve O\u27Connor, and Danny Fanning, as well as John Wolfe talking with the city licenser for alcohol, Don Greenhill about licensing, and Joe Lamb editor of the Morehead News, finally a small program is featured at the end involving an interview between Becky Smith and Larry Gooding, of the Flemingsburg Community Theatre, about a possible new theatre for them, compiled on September 30, 1982