Client processing is altered by novel myopathy-causing mutations in the HSP40 J domain

The misfolding and aggregation of proteins is often implicated in the development and progression of degenerative diseases. Heat shock proteins (HSPs), such as the ubiquitously expressed Type II Hsp40 molecular chaperone, DNAJB6, assist in protein folding and disaggregation. Historically, mutations within the DNAJB6 G/F domain have been associated with Limb-Girdle Muscular Dystrophy type 1D, now referred to as LGMDD1, a dominantly inherited degenerative disease. Recently, novel mutations within the J domain of DNAJB6 have been reported in patients with LGMDD1. Since novel myopathy-causing mutations in the Hsp40 J domain have yet to be characterized and both the function of DNAJB6 in skeletal muscle and the clients of this chaperone are unknown, we set out to assess the effect of these mutations on chaperone function using the genetically tractable yeast system. The essential yeast Type II Hsp40, Sis1, is homologous to DNAJB6 and is involved in the propagation of yeast prions. Using phenotypic, biochemical, and functional assays we found that homologous mutations in the Sis1 J domain differentially alter the processing of specific yeast prion strains, as well as a non-prion substrate. These data suggest that the newly-identified mutations in the J domain of DNAJB6 cause aberrant chaperone function that leads to the pathogenesis in LGMDD1

Metal-Poor Stars Observed with the Magellan Telescope. III. New Extremely and Ultra Metal-Poor Stars from SDSS/SEGUE and Insights on the Formation of Ultra Metal-Poor Stars

We report the discovery of one extremely metal-poor (EMP; [Fe/H]<-3) and one ultra metal-poor (UMP; [Fe/H]<-4) star selected from the SDSS/SEGUE survey. These stars were identified as EMP candidates based on their medium-resolution (R~2,000) spectra, and were followed-up with high-resolution (R~35,000) spectroscopy with the Magellan-Clay Telescope. Their derived chemical abundances exhibit good agreement with those of stars with similar metallicities. We also provide new insights on the formation of the UMP stars, based on comparison with a new set of theoretical models of supernovae nucleosynthesis. The models were matched with 20 UMP stars found in the literature, together with one of the program stars (SDSS J1204+1201), with [Fe/H]=-4.34. From fitting their abundances, we find that the supernovae progenitors, for stars where carbon and nitrogen are measured, had masses ranging from 20.5 M_sun to 28 M_sun and explosion energies from 0.3 to 0.9x10^51 erg. These results are highly sensitive to the carbon and nitrogen abundance determinations, which is one of the main drivers for future high-resolution follow-up of UMP candidates. In addition, we are able to reproduce the different CNO abundance patterns found in UMP stars with a single progenitor type, by varying its mass and explosion energy.Comment: 15 pages, 12 figures; accepted for publication in Ap

Evolved Gas Measurements Planned for the Lower Layers of the Gale Crater Mound with the Sample Analysis at Mars Instrument Suite

The lower mound strata of Gale Crater provide a diverse set of chemical environments for exploration by the varied tools of the Curiosity Rover of the Mars Science Laboratory (MSL) Mission. Orbital imaging and spectroscopy clearly reveal distinct layers of hydrated minerals, sulfates, and clays with abundant evidence of a variety of fluvial processes. The three instruments of the MSL Sample Analysis at aMars (SAM) investigation, the Quadrupole Mass Spectrometer (QMS), the Tunable Laser Spectrometer (TLS), and the Gas Chromatograph (GC) are designed to analyze either atmospheric gases or volatiles thermally evolved or chemically extracted from powdered rock or soil. The presence or absence of organic compounds in these layers is of great interest since such an in situ search for this type of record has not been successfully implemented since the mid-60s Viking GCMS experiments. However, regardless of the outcome of the analysis for organics, the abundance and isotopic composition of thermally evolved inorganic compounds should also provide a rich data set to complement the mineralogical and elemental information provided by other MSL instruments. In addition, these evolved gas analysis (EGA) experiments will help test sedimentary models proposed by Malin and Edgett (2000) and then further developed by Milliken et al (2010) for Gale Crater. In the SAM EGA experiments the evolution temperatures of H2O, CO2, SO2, O2, or other simple compounds as the samples are heated in a helium stream to 1000 C provides information on mineral types and their associations. The isotopic composition of O, H, C, and S can be precisely determined in several evolved compounds and compared with the present day atmosphere. Such SAM results might be able to test mineralogical evidence of changing sedimentary and alteration processes over an extended period of time. For example, Bibring et al (2006) have suggested such a major shift from early nonacidic to later acidic alteration. We will illustrate through a variety of evolved gas experiments implemented under SAM-like gas flow and temperature ramp conditions on terrestrial analog minerals on high fidelity Sam breadboards the type of chemical information we expect SAM to provide

Using whole-genome sequences of the LG/J and SM/J inbred mouse strains to prioritize quantitative trait genes and nucleotides

Background The laboratory mouse is the most commonly used model for studying variation in complex traits relevant to human disease. Here we present the whole-genome sequences of two inbred strains, LG/J and SM/J, which are frequently used to study variation in complex traits as diverse as aging, bone-growth, adiposity, maternal behavior, and methamphetamine sensitivity. Results We identified small nucleotide variants (SNVs) and structural variants (SVs) in the LG/J and SM/J strains relative to the reference genome and discovered novel variants in these two strains by comparing their sequences to other mouse genomes. We find that 39% of the LG/J and SM/J genomes are identical-by-descent (IBD). We characterized amino-acid changing mutations using three algorithms: LRT, PolyPhen-2 and SIFT. We also identified polymorphisms between LG/J and SM/J that fall in regulatory regions and highly informative transcription factor binding sites (TFBS). We intersected these functional predictions with quantitative trait loci (QTL) mapped in advanced intercrosses of these two strains. We find that QTL are both over-represented in non-IBD regions and highly enriched for variants predicted to have a functional impact. Variants in QTL associated with metabolic (231 QTL identified in an F16 generation) and developmental (41 QTL identified in an F34generation) traits were interrogated and we highlight candidate quantitative trait genes (QTG) and nucleotides (QTN) in a QTL on chr13 associated with variation in basal glucose levels and in a QTL on chr6 associated with variation in tibia length. Conclusions We show how integrating genomic sequence with QTL reduces the QTL search space and helps researchers prioritize candidate genes and nucleotides for experimental follow-up. Additionally, given the LG/J and SM/J phylogenetic context among inbred strains, these data contribute important information to the genomic landscape of the laboratory mouse

Monitoring, Restoration, and Source Water Protection: Canadian Community-Based Environmental Organizations’ Efforts towards Improving Aquatic Ecosystem Health

Published versionIn Canada, environmental monitoring has been the responsibility of government for decades; however, funding cutbacks have left many agencies unable to provide comprehensive coverage. This has stimulated a rise in community-based water monitoring (CBWM) organizations. These organizations, operating at multiple scales, have tasked themselves with monitoring aquatic ecosystems. Additionally, they often engage in restoration projects stemming from their monitoring work. Despite the growing abundance of CBWM organizations, there is uncertainty as to whether their activities lead to aquatic ecosystem benefits. A thematic analysis of photographic and qualitative interview data was employed to examine restoration projects conducted by five CBWM organizations, and the projects’ potential impact on source waters. Findings show that while they are conducting activities that show physical change, which is indicative of ecosystem improvement, examples of measurable responses within aquatic ecosystems remain rare. Monitoring, restoration, and source water protection processes are challenged by a lack of funding, capacity, and monitoring procedures. Funding, particularly, restricted the extent to which monitoring could be conducted and influenced project scope and scale. This leads to a lack of capacity to conduct large-scale restoration and rigorous scientific monitoring. Consequently, our findings highlight the issues with detecting effects of small-scale projects at the watershed scale

Disease-associated mutations within the yeast DNAJB6 homolog Sis1 slow conformer-specific substrate processing and can be corrected by the modulation of nucleotide exchange factors

Molecular chaperones, or heat shock proteins (HSPs), protect against the toxic misfolding and aggregation of proteins. As such, mutations or deficiencies within the chaperone network can lead to disease. Dominant mutations within DNAJB6 (Hsp40)-an Hsp70 co-chaperone-lead to a protein aggregation-linked myopathy termed Limb-Girdle Muscular Dystrophy Type D1 (LGMDD1). Here, we used the yeast prion model client in conjunction with in vitro chaperone activity assays to gain mechanistic insights into the molecular basis of LGMDD1. Here, we show how mutations analogous to those found in LGMDD1 affect Sis1 (a functional homolog of human DNAJB6) function by altering the structure of client protein aggregates, interfering with the Hsp70 ATPase cycle, dimerization and substrate processing; poisoning the function of wild-type protein. These results uncover the mechanisms through which LGMDD1-associated mutations alter chaperone activity, and provide insights relevant to potential therapeutic interventions