Mechanisms for quality control of misfolded transmembrane proteins

To prevent the accumulation of misfolded and aggregated proteins, the cell has developed a complex network of cellular quality control (QC) systems to recognize misfolded proteins and facilitate their refolding or degradation. The cell faces numerous obstacles when performing quality control on transmembrane proteins. Transmembrane proteins have domains on both sides of a membrane and QC systems in distinct compartments must coordinate to monitor the folding status of the protein. Additionally, transmembrane domains can have very complex organization and QC systems must be able to monitor the assembly of transmembrane domains in the membrane. In this review, we will discuss the QC systems involved in repair and degradation of misfolded transmembrane proteins. Also, we will elaborate on the factors that recognize folding defects of transmembrane domains and what happens when misfolded transmembrane proteins escape QC and aggregate

Dynamic Subunit Exchange and the Regulation of Microtubule Assembly by the Stress Response Protein Human αB Crystallin

BackgroundThe small heat shock protein (sHSP), human αB crystallin, forms large, polydisperse complexes that modulate the tubulin-microtubule equilibrium using a dynamic mechanism that is poorly understood. The interactive sequences in αB crystallin for tubulin are surface exposed, and correspond to interactive sites for the formation of αB crystallin complexes.Methodology/Principal FindingsThere is sequence homology between tubulin and the interactive domains in the β8-strand of the core domain and the C-terminal extension of αB crystallin. This study investigated the hypothesis that the formation of tubulin and αB crystallin quaternary structures was regulated through common interactive domains that alter the dynamics of their assembly. Size exclusion chromatography (SEC), SDS-PAGE, microtubule assembly assays, aggregation assays, multiple sequence alignment, and molecular modeling characterized the dynamic response of tubulin assembly to increasing concentrations of αB crystallin. Low molar ratios of αB crystallin∶tubulin were favorable for microtubule assembly and high molar ratios of αB crystallin∶tubulin were unfavorable for microtubule assembly. Interactions between αB crystallin and unassembled tubulin were observed using SEC and SDS-PAGE.Conclusions/SignificanceSubunits of αB crystallin that exchange dynamically with the αB crystallin complex can interact with tubulin subunits to regulate the equilibrium between tubulin and microtubules

On the Excitation and Formation of Circumstellar Fullerenes

We compare and analyze the Spitzer mid-infrared spectrum of three fullerene-rich planetary nebulae in the Milky Way and the Magellanic Clouds; Tc1, SMP SMC16, and SMP LMC56. The three planetary nebulae share many spectroscopic similarities. The strongest circumstellar emission bands correspond to the infrared active vibrational modes of the fullerene species C60 and little or no emission is present from Polycyclic Aromatic Hydrocarbons (PAHs). The strength of the fullerene bands in the three planetary nebulae is very similar, while the ratio of the [NeIII]15.5um/[NeII]12.8um fine structure lines, an indicator of the strength of the radiation field, is markedly different. This raises questions about their excitation mechanism and we compare the fullerene emission to fluorescent and thermal models. In addition, the spectra show other interesting and common features, most notably in the 6-9um region, where a broad plateau with substructure dominates the emission. These features have previously been associated with mixtures of aromatic/aliphatic hydrocarbon solids. We hypothesize on the origin of this band, which is likely related to the fullerene formation mechanism, and compare it with modeled Hydrogenated Amorphous Carbon that present emission in this region.Comment: 13 pages, 2 tables, 7 figures, Accepted for publication in Ap

Restoration of R117H CFTR folding and function in human airway cells through combination treatment with VX-809 and VX-770

Cystic fibrosis (CF) is a lethal recessive genetic disease caused primarily by the F508del mutation in the CF transmembrane conductance regulator (CFTR). The potentiator VX-770 was the first CFTR modulator approved by the FDA for treatment of CF patients with the gating mutation G551D. Orkambi is a drug containing VX-770 and corrector VX809 and is approved for treatment of CF patients homozygous for F508del, which has folding and gating defects. At least 30% of CF patients are heterozygous for the F508del mutation with the other allele encoding for one of many different rare CFTR mutations. Treatment of heterozygous F508del patients with VX-809 and VX-770 has had limited success, so it is important to identify heterozygous patients that respond to CFTR modulator therapy. R117H is a more prevalent rare mutation found in over 2,000 CF patients. In this study we investigated the effectiveness of VX-809/VX-770 therapy on restoring CFTR function in human bronchial epithelial (HBE) cells from R117H/F508del CF patients. We found that VX-809 stimulated more CFTR activity in R117H/F508del HBEs than in F508del/F508del HBEs. R117H expressed exclusively in immortalized HBEs exhibited a folding defect, was retained in the ER, and degraded prematurely. VX-809 corrected the R117H folding defect and restored channel function. Because R117 is involved in ion conductance, VX-770 acted additively with VX-809 to restore CFTR function in chronically treated R117H/F508del cells. Although treatment of R117H patients with VX-770 has been approved, our studies indicate that Orkambi may be more beneficial for rescue of CFTR function in these patients

Starburst and cirrus models for submillimeter galaxies

We present radiative transfer models for submillimeter galaxies with spectroscopic redshifts and mid-infrared spectroscopy from Spitzer/IRS and analyze available Spitzer/MIPS 24, 70 and 160mu data. We use two types of starburst models, a cirrus model and a model for the emission of an AGN torus in order to investigate the nature of these objects. We find that for three of the objects (25%) cirrus emission alone can account for the mid-infrared spectrum and the MIPS and submillimeter data. For the remaining objects we find that we need a combination of starburst and cirrus in order to fit simultaneously the multi--wavelength data. We find that the typical submillimeter galaxy has comparable luminosity in the starburst (median L=10^12.5 Lo) and cirrus (median L=10^12.4 Lo) components. This could arise if the galaxies have been forming stars continuously for the last 250Myr with the star formation occurring in the last 5Myr being shrouded by high-optical-depth molecular cloud dust, whereas the rest of the starlight is attenuated by diffuse dust or cirrus with an Av of about 1mag.Comment: 9 pages, AA accepte

Spitzer reveals what's behind Orion's Bar

We present Spitzer Space Telescope observations of 11 regions SE of the Bright Bar in the Orion Nebula, along a radial from the exciting star theta1OriC, extending from 2.6 to 12.1'. Our Cycle 5 programme obtained deep spectra with matching IRS short-high (SH) and long-high (LH) aperture grid patterns. Most previous IR missions observed only the inner few arcmin. Orion is the benchmark for studies of the ISM particularly for elemental abundances. Spitzer observations provide a unique perspective on the Ne and S abundances by virtue of observing the dominant ionization states of Ne (Ne+, Ne++) and S (S++, S3+) in Orion and H II regions in general. The Ne/H abundance ratio is especially well determined, with a value of (1.01+/-0.08)E-4. We obtained corresponding new ground-based spectra at CTIO. These optical data are used to estimate the electron temperature, electron density, optical extinction, and the S+/S++ ratio at each of our Spitzer positions. That permits an adjustment for the total gas-phase S abundance because no S+ line is observed by Spitzer. The gas-phase S/H abundance ratio is (7.68+/-0.30)E-6. The Ne/S abundance ratio may be determined even when the weaker hydrogen line, H(7-6) here, is not measured. The mean value, adjusted for the optical S+/S++ ratio, is Ne/S = 13.0+/-0.6. We derive the electron density versus distance from theta1OriC for [S III] and [S II]. Both distributions are for the most part decreasing with increasing distance. A dramatic find is the presence of high-ionization Ne++ all the way to the outer optical boundary ~12' from theta1OriC. This IR result is robust, whereas the optical evidence from observations of high-ionization species (e.g. O++) at the outer optical boundary suffers uncertainty because of scattering of emission from the much brighter inner Huygens Region.Comment: 60 pages, 16 figures, 10 tables. MNRAS accepte

MBM 12: young protoplanetary discs at high galactic latitude

(abridged) We present Spitzer infrared observations to constrain disc and dust evolution in young T Tauri stars in MBM 12, a star-forming cloud at high latitude with an age of 2 Myr and a distance of 275 pc. The region contains 12 T Tauri systems, with primary spectral types between K3 and M6; 5 are weak-line and the rest classical T Tauri stars. We first use MIPS and literature photometry to compile spectral energy distributions for each of the 12 members in MBM 12, and derive their IR excesses. The IRS spectra are analysed with the newly developed two-layer temperature distribution (TLTD) spectral decomposition method. For the 7 T Tauri stars with a detected IR excess, we analyse their solid-state features to derive dust properties such as mass-averaged grain size, composition and crystallinity. We find a spatial gradient in the forsterite to enstatite range, with more enstatite present in the warmer regions. The fact that we see a radial dependence of the dust properties indicates that radial mixing is not very efficient in the discs of these young T Tauri stars. The SED analysis shows that the discs in MBM 12, in general, undergo rapid inner disc clearing, while the binary sources have faster discevolution. The dust grains seem to evolve independently from the stellar properties, but are mildly related to disc properties such as flaring and accretion rates.Comment: 14 pages, accepted by Astronomy and Astrophysic

A far-infrared spectroscopic survey of intermediate redshift (ultra) luminous infrared galaxies

We present Herschel far-IR photometry and spectroscopy as well as ground-based CO observations of an intermediate redshift (0.21 _ z _ 0.88) sample of Herschel-selected (ultra)-luminous infrared galaxies (LIR >1011.5 L_). With these measurements, we trace the dust continuum, far-IR atomic line emission, in particular [C ii] 157.7μm, as well as the molecular gas of z ∼ 0.3 luminous and ultraluminous infrared galaxies (LIRGs and ULIRGs) and perform a detailed investigation of the interstellar medium of the population. We find that the majority of Herschel-selected intermediate redshift (U)LIRGs have LCii/LFIR ratios that are a factor of about 10 higher than that of local ULIRGs and comparable to that of local normal and high-z star-forming galaxies. Using our sample to bridge local and high-z [C ii] observations, we find that the majority of galaxies at all redshifts and all luminosities follow an LCii−LFIR relation with a slope of unity, from which local ULIRGs and high-z active-galactic-nucleus-dominated sources are clear outliers.We also confirm that the strong anti-correlation between the LCii/LFIR ratio and the far-IR color L60/L100 observed in the local universe holds over a broad range of redshifts and luminosities, in the sense that warmer sources exhibit lower LC ii/LFIR at any epoch. Intermediate redshift ULIRGs are also characterized by large molecular gas reservoirs and by lower star formation efficiencies compared to that of local ULIRGs. The high LC ii/LFIR ratios, the moderate star formation efficiencies (LIR/L _COor LIR/MH2 ), and the relatively low dust temperatures of our sample (which are also common characteristics of high-z star-forming galaxies with ULIRG-like luminosities) indicate that the evolution of the physical properties of (U)LIRGs between the present day and z > 1 is already significant by z ∼ 0.3

From CFTR biology toward combinatorial pharmacotherapy:expanded classification of cystic fibrosis mutations

More than 2000 mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) have been described that confer a range of molecular cell biological and functional phenotypes. Most of these mutations lead to compromised anion conductance at the apical plasma membrane of secretory epithelia and cause cystic fibrosis (CF) with variable disease severity. Based on the molecular phenotypic complexity of CFTR mutants and their susceptibility to pharmacotherapy, it has been recognized that mutations may impose combinatorial defects in CFTR channel biology. This notion led to the conclusion that the combination of pharmacotherapies addressing single defects (e.g., transcription, translation, folding, and/or gating) may show improved clinical benefit over available low-efficacy monotherapies. Indeed, recent phase 3 clinical trials combining ivacaftor (a gating potentiator) and lumacaftor (a folding corrector) have proven efficacious in CF patients harboring the most common mutation (deletion of residue F508, ΔF508, or Phe508del). This drug combination was recently approved by the U.S. Food and Drug Administration for patients homozygous for ΔF508. Emerging studies of the structural, cell biological, and functional defects caused by rare mutations provide a new framework that reveals a mixture of deficiencies in different CFTR alleles. Establishment of a set of combinatorial categories of the previously defined basic defects in CF alleles will aid the design of even more efficacious therapeutic interventions for CF patients