Proteasome storage granules protect proteasomes from autophagic degradation upon carbon starvation

26S proteasome abundance is tightly regulated at multiple levels, including the elimination of excess or inactive particles by autophagy. In yeast, this proteaphagy occurs upon nitrogen starvation but not carbon starvation, which instead stimulates the rapid sequestration of proteasomes into cytoplasmic puncta termed proteasome storage granules (PSGs). Here, we show that PSGs help protect proteasomes from autophagic degradation. Both the core protease and regulatory particle sub-complexes are sequestered separately into PSGs via pathways dependent on the accessory proteins Blm10 and Spg5, respectively. Modulating PSG formation, either by perturbing cellular energy status or pH, or by genetically eliminating factors required for granule assembly, not only influences the rate of proteasome degradation, but also impacts cell viability upon recovery from carbon starvation. PSG formation and concomitant protection against proteaphagy also occurs in Arabidopsis, suggesting that PSGs represent an evolutionarily conserved cache of proteasomes that can be rapidly re-mobilized based on energy availability

Autophagic Turnover of Inactive 26S Proteasomes in Yeast Is Directed by the Ubiquitin Receptor Cue5 and the Hsp42 Chaperone

Highlights The yeast 26S proteasome is degraded by Atg8-mediated autophagy Nitrogen starvation and inactivation stimulate proteaphagy via distinct pathways Proteasome inhibition is accompanied by extensive ubiquitylation of the complex Proteaphagy engages the Cue5 autophagy receptor and the Hsp42 chaperone Summary The autophagic clearance of 26S proteasomes (proteaphagy) is an important homeostatic mechanism within the ubiquitin system that modulates proteolytic capacity and eliminates damaged particles. Here, we define two proteaphagy routes in yeast that respond to either nitrogen starvation or particle inactivation. Whereas the core autophagic machineries required for Atg8 lipidation and vesiculation are essential for both routes, the upstream Atg1 kinase participates only in starvation-induced proteaphagy. Following inactivation, 26S proteasomes become extensively modified with ubiquitin. Although prior studies with Arabidopsis implicated RPN10 in tethering ubiquitylated proteasomes to ATG8 lining the autophagic membranes, yeast proteaphagy employs the evolutionarily distinct receptor Cue5, which simultaneously binds ubiquitin and Atg8. Proteaphagy of inactivated proteasomes also requires the oligomeric Hsp42 chaperone, suggesting that ubiquitylated proteasomes are directed by Hsp42 to insoluble protein deposit (IPOD)-type structures before encapsulation. Together, Cue5 and Hsp42 provide a quality control checkpoint in yeast directed at recycling dysfunctional 26S proteasomes

Purification of 26S Proteasomes and Their Subcomplexes from Plants

The 26S proteasome is a highly dynamic, multisubunit, ATP-dependent protease that plays a central role in cellular housekeeping and many aspects of plant growth and development by degrading aberrant polypeptides and key cellular regulators that are first modified by ubiquitin. Although the 26S proteasome was originally enriched from plants over 30 years ago, only recently have significant advances been made in our ability to isolate and study the plant particle. Here, we describe two robust methods for purifying the 26S proteasome and its subcomplexes from Arabidopsis thaliana; one that involves conventional chromatography techniques to isolate the complex from wild-type plants, and another that employs the genetic replacement of individual subunits with epitope-tagged variants combined with affinity purification. In addition to these purification protocols, we describe methods commonly used to analyze the activity and composition of the complex

DESIGNING WETLAND CONSERVATION STRATEGIES UNDER CLIMATE CHANGE

A methodology for evaluating public wetlands conservation investments that considers climate change is developed and applied to Virginia's Elizabeth River watershed. A revised cellular automaton (CA) model is applied to project future land use change. Discrete stochastic sequential programming (DSSP) is used to model a parcel-based discrete-time decision process.Environmental Economics and Policy,

A massive warm baryonic halo in the Coma cluster

Several deep PSPC observations of the Coma cluster reveal a very large-scale halo of soft X-ray emission, substantially in excess of the well known radiation from the hot intra-cluster medium. The excess emission, previously reported in the central region of the cluster using lower-sensitivity EUVE and ROSAT data, is now evident out to a radius of 2.6 Mpc, demonstrating that the soft excess radiation from clusters is a phenomenon of cosmological significance. The X-ray spectrum at these large radii cannot be modeled non-thermally, but is consistent with the original scenario of thermal emission from warm gas at ~ 10^6 K. The mass of the warm gas is on par with that of the hot X-ray emitting plasma, and significantly more massive if the warm gas resides in low-density filamentary structures. Thus the data lend vital support to current theories of cosmic evolution, which predict that at low redshift \~30-40 % of the baryons reside in warm filaments converging at clusters of galaxies.Comment: Astrophysical Journal, in pres

Correlated X-ray and Optical Variability in Mkn 509

We present results of a 3 year monitoring campaign of the Seyfert 1 galaxy Markarian 509, using X-ray data from the Rossi X-ray Timing Explorer (RXTE) and optical data taken by the SMARTS consortium. Both light curves show significant variations, and are strongly correlated with the optical flux leading the X-ray flux by 15 days. The X-ray power spectrum shows a steep high-frequency slope of -2.0, breaking to a slope of -1.0 at at timescale of 34 days. The lag from optical to X-ray emission is most likely caused by variations in the accretion disk propagating inward.Comment: 13 pages, 3 figures. Accepted for publication in the Astrophysical Journa

Integrated stratigraphy of the Kimmeridge Clay Formation (Upper Jurassic) based on exposures and boreholes in south Dorset, UK

For the purposes of a high-resolution multi-disciplinary study of the Upper Jurassic Kimmeridge Clay Formation, two boreholes were drilled at Swanworth Quarry and one at Metherhills, south Dorset, UK. Together, the cores represent the first complete section through the entire formation close to the type section. We present graphic logs that record the stratigraphy of the cores, and outline the complementary geophysical and analytical data sets (gamma ray, magnetic susceptibility, total organic carbon, carbonate, [delta]13Corg). Of particular note are the new borehole data from the lowermost part of the formation which does not crop out in the type area. Detailed logs are available for download from the Kimmeridge Drilling Project web-site at http://kimmeridge.earth.ox.ac.uk/. Of further interest is a mid-eudoxus Zone positive shift in the [delta]13Corg record, a feature that is also registered in Tethyan carbonate successions, suggesting that it is a regional event and may therefore be useful for correlation. The lithostratigraphy of the cores has been precisely correlated with the nearby cliff section, which has also been examined and re-described. Magnetic-susceptibility and spectral gamma-ray measurements were made at a regular spacing through the succession, and facilitate core-to-exposure correlation. The strata of the exposure and core have been subdivided into four main mudrock lithological types: (a) medium-dark–dark-grey marl; (b) medium-dark–dark grey–greenish black shale; (c) dark-grey–olive-black laminated shale; (d) greyish-black–brownish-black mudstone. The sections also contain subordinate amounts of siltstone, limestone and dolostone. Comparison of the type section with the cores reveals slight lithological variation and notable thickness differences between the coeval strata. The proximity of the boreholes and different parts of the type section to the Purbeck–Isle of Wight Disturbance is proposed as a likely control on the thickness changes

RETROCAM: A Versatile Optical Imager for Synoptic Studies

We present RETROCAM, an auxiliary CCD camera that can be rapidly inserted into the optical beam of the MDM 2.4m telescope. The speed and ease of reconfiguring the telescope to use the imager and a straightforward user interface permit the camera to be used during the course of other observing programs. This in turn encourages RETROCAM's use for a variety of monitoring projects.Comment: 6 pages, 6 figures, Accepted by A

Performance, Politics and Media: How the 2010 British General Election leadership debates generated ‘talk’ amongst the electorate.

During the British General Election 2010 a major innovation was introduced in part to improve engagement: a series of three live televised leadership debates took place where the leader of each of the three main parties, Labour, Liberal Democrat and Conservative, answered questions posed by members of the public and subsequently debated issues pertinent to the questions. In this study we consider these potentially ground breaking debates as the kind of event that was likely to generate discussion. We investigate various aspects of the ‘talk’ that emerged as a result of watching the debates. As an exploratory study concerned with situated accounts of the participants experiences we take an interpretive perspective. In this paper we outline the meta-narratives (of talk) associated with the viewing of the leadership debates that were identified, concluding our analysis by suggesting that putting a live debate on television and promoting and positioning it as a major innovation is likely to mean that is how the audience will make sense of it – as a media event