Extra N-Terminal Residues Have a Profound Effect on the Aggregation Properties of the Potential Yeast Prion Protein Mca1

The metacaspase Mca1 from Saccharomyces cerevisiae displays a Q/N-rich region at its N-terminus reminiscent of yeast prion proteins. In this study, we show that the ability of Mca1 to form insoluble aggregates is modulated by a peptide stretch preceding its putative prion-forming domain. Based on its genomic locus, three potential translational start sites of Mca1 can give rise to two slightly different long Mca1 proteins or a short version, Mca1451/453 and Mca1432, respectively, although under normal physiological conditions Mca1432 is the predominant form expressed. All Mca1 variants exhibit the Q/N-rich regions, while only the long variants Mca1451/453 share an extra stretch of 19 amino acids at their N-terminal end. Strikingly, only long versions of Mca1 but not Mca1432 revealed pronounced aggregation in vivo and displayed prion-like properties when fused to the C-terminal domain of Sup35 suggesting that the N-terminal peptide element promotes the conformational switch of Mca1 protein into an insoluble state. Transfer of the 19 N-terminal amino acid stretch of Mca1451 to the N-terminus of firefly luciferase resulted in increased aggregation of luciferase, suggesting a protein destabilizing function of the peptide element. We conclude that the aggregation propensity of the potential yeast prion protein Mca1 in vivo is strongly accelerated by a short peptide segment preceding its Q/N-rich region and we speculate that such a conformational switch might occur in vivo via the usage of alternative translational start sites

Exposure age and climate controls on weathering in deglaciated watersheds of western Greenland

Fine-grained sediments deposited by retreating glaciers weather faster than the global average and this weathering can impact the global carbon cycle and oceanic fluxes of nutrients and radiogenic isotopes. Much work has focused on subglacial and proglacial weathering of continental ice sheets, but little is known about weathering and resulting fluxes from deglacial watersheds, which are disconnected from the ice sheets and discharge only annual precipitation and permafrost melt. We investigate the effects of exposure age and precipitation on weathering intensity in four deglacial watersheds on Greenland that form a transect from the coast near Sisimiut toward the Greenland Ice Sheet (GrIS) near Kangerlussuaq based on evaluations of major ion compositions, Sr isotope ratios, and mineral saturation states of waters and sediments. The transect is underlain by Archean orthogneiss and is characterized by gradients in moraine ages (∼7.5–8.0 ky inland to ∼10 ky at the coast) and water balance (−150 mm/yr inland to +150 mm/yr at the coast). Anion compositions are generally dominated by HCO3, but SO4 becomes increasingly important toward the coast, reflecting a switch from trace carbonate dissolution to sulfide mineral oxidation. Coastal watersheds have a higher proportion of dissolved silica, higher Na/Cl, Si/Ca, and lower Ca/Sr ratios than inland watersheds, indicating an increase in the relative proportion of silicate weathering and an increase in the extent of weathering toward the coast. More extensive weathering near the coast is also apparent in differences in the 87Sr/86Sr ratios of stream water and bedload (Δ87Sr/86Sr), which decreases from 0.017 inland to 0.005 at the coast, and in increased saturation states relative to amorphous SiO2 and quartz. The steep weathering gradient from inland to coastal watersheds reflects enhanced weathering compared to that expected from the 2 to 3 ky difference in exposure age caused by elevated coastal precipitation. The gradient of weathering with exposure age, water budget and distance from the ice sheet indicates that oceanic and atmospheric fluxes will change as continental glaciers retreat, precipitation patterns across the deglacial region readjust, and the relative proportion of deglacial to proglacial runoff increases

The soil geochemistry in the Beardmore Glacier Region, Antarctica: Implications for terrestrial ecosystem history

Although most models suggest continental Antarctica was covered by ice during the Last Glacial Maximum (LGM) it has been speculated that endemic species of soil invertebrates could have survived the Pleistocene at high elevation habitats protruding above the ice sheets. We analyzed a series of soil samples from different elevations at three locations along the Beardmore Glacier in the Transantarctic Mountains (in order of increasing elevation): Ebony Ridge (ER), Cloudmaker (CM), and Meyer Desert (MD). Geochemical analyses show the MD soils, which were exposed during the LGM, were the least weathered compared to lower elevations, and also had the highest total dissolved solids (TDS). MD soils are dominated by nitrate salts (NO₃/Cl ratios >10) that can be observed in SEM images. High δ¹⁷O and δ¹⁸O values of the nitrate indicate that its source is solely of atmospheric origin. It is suggested that nitrate concentrations in the soil may be utilized to determine a relative “wetting age” to better assess invertebrate habitat suitability. The highest elevation sites at MD have been exposed and accumulating salts for the longest times, and because of the salt accumulations, they were not suitable as invertebrate refugia during the LGM

Photon detector system timing performance in the DUNE 35-ton prototype liquid argon time projection chamber

The 35-ton prototype for the Deep Underground Neutrino Experiment far detector was a single-phase liquid argon time projection chamber with an integrated photon detector system, all situated inside a membrane cryostat. The detector took cosmic-ray data for six weeks during the period of February 1, 2016 to March 12, 2016. The performance of the photon detection system was checked with these data. An installed photon detector was demonstrated to measure the arrival times of cosmic-ray muons with a resolution better than 32 ns, limited by the timing of the trigger system. A measurement of the timing resolution using closely-spaced calibration pulses yielded a resolution of 15 ns for pulses at a level of 6 photo-electrons. Scintillation light from cosmic-ray muons was observed to be attenuated with increasing distance with a characteristic length of 155 ± 28 cm

Fermilab Main Injector Beam Position Monitor Upgrade

An upgrade of the Beam Position Monitor (BPM) signal processing and data acquisition system for the Fermilab Main Injector is described. The Main Injector is a fast cycling synchrotron that accelerates protons or antiprotons from 8 to 150 GeV. Each Main Injector cycle can have a totally different magnet ramp, RF frequency configuration, beam bunch structure, and injection/extraction pattern from the previous cycle. The new BPM system provides the capabilities and flexibility required by the dynamic and complex machine operations. The system offers measurement capability in the 2.5 MHz and 53 MHz channels to detect the range of bunch structures for protons and antiprotons in both wideband (turn-by-turn) and narrowband (closed-orbit) modes. The new BPM read-out system is based on the digital receiver concept and is highly configurable, allowing the signal processing of nearly all Main Injector beam conditions, including the detection of individual batches. An overview of the BPM system in the Main Injector operating environment, some technology details and first beam measurements are presented

Protein folding on the ribosome studied using NMR spectroscopy

NMR spectroscopy is a powerful tool for the investigation of protein folding and misfolding, providing a characterization of molecular structure, dynamics and exchange processes, across a very wide range of timescales and with near atomic resolution. In recent years NMR methods have also been developed to study protein folding as it might occur within the cell, in a de novo manner, by observing the folding of nascent polypeptides in the process of emerging from the ribosome during synthesis. Despite the 2.3 MDa molecular weight of the bacterial 70S ribosome, many nascent polypeptides, and some ribosomal proteins, have sufficient local flexibility that sharp resonances may be observed in solution-state NMR spectra. In providing information on dynamic regions of the structure, NMR spectroscopy is therefore highly complementary to alternative methods such as X-ray crystallography and cryo-electron microscopy, which have successfully characterized the rigid core of the ribosome particle. However, the low working concentrations and limited sample stability associated with ribosome-nascent chain complexes means that such studies still present significant technical challenges to the NMR spectroscopist. This review will discuss the progress that has been made in this area, surveying all NMR studies that have been published to date, and with a particular focus on strategies for improving experimental sensitivity

Heterologous Protein Expression Is Enhanced by Harmonizing the Codon Usage Frequencies of the Target Gene with those of the Expression Host

Synonymous codon replacement can change protein structure and function, indicating that protein structure depends on DNA sequence. During heterologous protein expression, low expression or formation of insoluble aggregates may be attributable to differences in synonymous codon usage between expression and natural hosts. This discordance may be particularly important during translation of the domain boundaries (link/end segments) that separate elements of higher ordered structure. Within such regions, ribosomal progression slows as the ribosome encounters clusters of infrequently used codons that preferentially encode a subset of amino acids. To replicate the modulation of such localized translation rates during heterologous expression, we used known relationships between codon usage frequencies and secondary protein structure to develop an algorithm (“codon harmonization”) for identifying regions of slowly translated mRNA that are putatively associated with link/end segments. It then recommends synonymous replacement codons having usage frequencies in the heterologous expression host that are less than or equal to the usage frequencies of native codons in the native expression host. For protein regions other than these putative link/end segments, it recommends synonymous substitutions with codons having usage frequencies matched as nearly as possible to the native expression system. Previous application of this algorithm facilitated E. coli expression, manufacture and testing of two Plasmodium falciparum vaccine candidates. Here we describe the algorithm in detail and apply it to E. coli expression of three additional P. falciparum proteins. Expression of the “recoded” genes exceeded that of the native genes by 4- to 1,000-fold, representing levels suitable for vaccine manufacture. The proteins were soluble and reacted with a variety of functional conformation-specific mAbs suggesting that they were folded properly and had assumed native conformation. Codon harmonization may further provide a general strategy for improving the expression of soluble functional proteins during heterologous expression in hosts other than E. coli

Mu2e Technical Design Report

The Mu2e experiment at Fermilab will search for charged lepton flavor violation via the coherent conversion process mu- N --> e- N with a sensitivity approximately four orders of magnitude better than the current world's best limits for this process. The experiment's sensitivity offers discovery potential over a wide array of new physics models and probes mass scales well beyond the reach of the LHC. We describe herein the preliminary design of the proposed Mu2e experiment. This document was created in partial fulfillment of the requirements necessary to obtain DOE CD-2 approval.Comment: compressed file, 888 pages, 621 figures, 126 tables; full resolution available at http://mu2e.fnal.gov; corrected typo in background summary, Table 3.

First measurement of muon-neutrino disappearance in NOvA

This paper reports the first measurement using the NOvA detectors of νμ disappearance in a νμ beam. The analysis uses a 14 kton-equivalent exposure of 2.74×1020 protons-on-target from the Fermilab NuMI beam. Assuming the normal neutrino mass hierarchy, we measure Δm232=(2.52+0.20−0.18)×10−3  eV2 and sin2θ23 in the range 0.38–0.65, both at the 68% confidence level, with two statistically degenerate best-fit points at sin2θ23=0.43 and 0.60. Results for the inverted mass hierarchy are also presented