Altered sterol metabolism in budding yeast affects mitochondrial iron–sulfur (Fe-S) cluster synthesis

Ergosterol synthesis is essential for cellular growth and viability of the budding yeast Saccharomyces cerevisiae, and intracellular sterol distribution and homeostasis are therefore highly regulated in this species. Erg25 is an iron-containing C4-methyl sterol oxidase that contributes to the conversion of 4,4-dimethylzymosterol to zymosterol, a precursor of ergosterol. The ERG29 gene encodes an endoplasmic reticulum (ER)-associated protein, and here we identified a role for Erg29 in the methyl sterol oxidase step of ergosterol synthesis. ERG29 deletion resulted in lethality in respiring cells, but respiration-incompetent (Rho- or Rho0) cells survived, suggesting that Erg29 loss leads to accumulation of oxidized sterol metabolites that affect cell viability. Down-regulation of ERG29 expression in Δerg29 cells indeed led to accumulation of methyl sterol metabolites, resulting in increased mitochondrial oxidants and a decreased ability of mitochondria to synthesize iron-sulfur (Fe-S) clusters due to reduced levels of Yfh1, the mammalian frataxin homolog, which is involved in mitochondrial iron metabolism. Using a high-copy genomic library, we identified suppressor genes that permitted growth of Δerg29 cells on respiratory substrates, and these included genes encoding the mitochondrial proteins Yfh1, Mmt1, Mmt2, and Pet20, which reversed all phenotypes associated with loss of ERG29 Of note, loss of Erg25 also resulted in accumulation of methyl sterol metabolites and also increased mitochondrial oxidants and degradation of Yfh1. We propose that accumulation of toxic intermediates of the methyl sterol oxidase reaction increases mitochondrial oxidants, which affect Yfh1 protein stability. These results indicate an interaction between sterols generated by ER proteins and mitochondrial iron metabolism

Charm Cross Sections for the Tevatron Run II

We present a calculation of the D^{*+}, D^+ and D^0 meson single inclusive production cross section for the Tevatron Run II. We use the FONLL approach in perturbative QCD, which, besides including the known next-to-leading order corrections, also provides for the resummation at the next-to-leading logarithmic level of terms enhanced at large p_T by powers of log(p_T/m), where m is the charm mass and p_T is its transverse momentum. Non-perturbative effects in charm hadronization are extracted, in moment space, from recent ALEPH data for D^* fragmentation in e^+e^- collisions.Comment: 11 pages, 5 figures, LaTe

Secondary ionization and heating by fast electrons

We examine the fate of fast electrons (with energies E>10 eV) in a thermal gas of primordial composition. To follow their interactions with the background gas, we construct a Monte Carlo model that includes: (1) electron-electron scattering (which transforms the electron kinetic energy into heat), (2) collisional ionization of hydrogen and helium (which produces secondary electrons that themselves scatter through the medium), and (3) collisional excitation (which produces secondary photons, whose fates we also follow approximately). For the last process, we explicitly include all transitions to upper levels n<=4, together with a well-motivated extrapolation to higher levels. In all cases, we use recent calculated cross-sections at E<1 keV and the Bethe approximation to extrapolate to higher energies. We compute the fractions of energy deposited as heat, ionization (tracking HI and the helium species separately), and excitation (tracking HI Lyman-alpha separately) under a broad range of conditions appropriate to the intergalactic medium. The energy deposition fractions depend on both the background ionized fraction and the electron energy but are nearly independent of the background density. We find good agreement with some, but not all, previous calculations at high energies. Electronic tables of our results are available on request.Comment: 10 pages, 6 figures, submitted to MNRA

Differences in leukocyte profile, gene expression, and metabolite status of dairy cows with or without sole ulcers

peer-reviewedSole ulcers are one of the most severe pathologies causing lameness in dairy cows and are associated with abnormal behavior and impaired production performance. However, little is known about how or whether lameness caused by sole ulcers affects the cow systemically. This study compared hematology profile, leukocyte gene expression, and physiological responses [metabolite, cortisol, the endogenous steroid hormone dehydroepiandrosterone (DHEA), and haptoglobin concentrations] of cows with sole ulcers and healthy cows. Twelve clinically lame cows (lame) were identified as having at least one sole ulcer and no other disorder, and matched with a cow that had good locomotion and no disorders (sound), using days in milk, liveweight, body condition score, and diet. Blood samples were taken from all 24 cows within 24 h of sole ulcer diagnosis. Leukocyte counts were obtained using an automated cell counter, cortisol and DHEA concentration by ELISA, and plasma haptoglobin, urea, total protein, creatine kinase, and glucose were analyzed on an Olympus analyzer. Expression of 16 genes associated with lameness or stress were estimated using reverse transcription-PCR. Data were analyzed using the MIXED procedure in SAS software (version 9.3; SAS Institute Inc., Cary, NC). Lame cows had a higher neutrophil percentage, a numerically lower lymphocyte percentage, and tended to have a higher neutrophil:lymphocyte ratio than sound cows. Serum cortisol and DHEA concentrations were higher in lame than in sound cows. Lame cows also tended to have higher haptoglobin and glucose levels than sound, as well as higher protein yet lower urea levels. Sound cows tended to have higher relative expression of the gene coding for colony-stimulating factor 2 than lame, but in all other cases where differences were detected in cytokine gene expression (IL-1α, IL-1β, CXCL8, and IL-10), relative gene expression in sound cows tended to be, or was, lower than in lame. Relative expression of MMP-13, GR-α, Fas, haptoglobin, and CD62L were, or tended to be, higher in lame than sound cows. A high neutrophil:lymphocyte ratio in combination with higher cortisol levels in cows with ulcers is indicative of physiological stress. Moreover, increased DHEA and a higher cortisol:DHEA ratio, as well as a tendency for higher haptoglobin levels and increased haptoglobin mRNA expression, are indicative of systemic inflammation. Increased cytokine mRNA expression indicates activation of the immune system compared with healthy cows. Increased expression of MMP-13 mRNA has been found in cows with impaired locomotion and thus could be implicated in development of claw horn disorders.This study was funded by a Marie Curie Intra-European Fellowship (FP7-People 2009-IEF; grant agreement number: 252611) to Keelin O'Driscoll

Cpd-1 Null Mice Display a Subtle Neurological Phenotype

CPD1 (also known as ANP32-E) belongs to a family of evolutionarily conserved acidic proteins with leucine rich repeats implicated in a variety of cellular processes regulating gene expression, vesicular trafficking, intracellular signaling and apoptosis. Because of its spatiotemporal expression pattern, CPD1 has been proposed to play an important role in brain morphogenesis and synaptic development.We have generated CPD1 knock-out mice that we have subsequently characterized. These mice are viable and fertile. However, they display a subtle neurological clasping phenotype and mild motor deficits.CPD1 is not essential for normal development; however, it appears to play a role in the regulation of fine motor functions. The minimal phenotype suggests compensatory biological mechanisms

LEM2 recruits CHMP7 for ESCRT-mediated nuclear envelope closure in fission yeast and human cells

Endosomal sorting complexes required for transport III (ESCRT-III) proteins have been implicated in sealing the nuclear envelope in mammals, spindle pole body dynamics in fission yeast, and surveillance of defective nuclear pore complexes in budding yeast. Here, we report that Lem2p (LEM2), a member of the LEM (Lap2-Emerin-Man1) family of inner nuclear membrane proteins, and the ESCRT-II/ESCRT-III hybrid protein Cmp7p (CHMP7), work together to recruit additional ESCRT-III proteins to holes in the nuclear membrane. In Schizosaccharomyces pombe, deletion of the ATPase vps4 leads to severe defects in nuclear morphology and integrity. These phenotypes are suppressed by loss-of-function mutations that arise spontaneously in lem2 or cmp7, implying that these proteins may function upstream in the same pathway. Building on these genetic interactions, we explored the role of LEM2 during nuclear envelope reformation in human cells. We found that CHMP7 and LEM2 enrich at the same region of the chromatin disk periphery during this window of cell division and that CHMP7 can bind directly to the C-terminal domain of LEM2 in vitro. We further found that, during nuclear envelope formation, recruitment of the ESCRT factors CHMP7, CHMP2A, and IST1/CHMP8 all depend on LEM2 in human cells. We conclude that Lem2p/LEM2 is a conserved nuclear site-specific adaptor that recruits Cmp7p/CHMP7 and downstream ESCRT factors to the nuclear envelope

General analysis of signals with two leptons and missing energy at the Large Hadron Collider

A signal of two leptons and missing energy is challenging to analyze at the Large Hadron Collider (LHC) since it offers only few kinematical handles. This signature generally arises from pair production of heavy charged particles which each decay into a lepton and a weakly interacting stable particle. Here this class of processes is analyzed with minimal model assumptions by considering all possible combinations of spin 0, 1/2 or 1, and of weak iso-singlets, -doublets or -triplets for the new particles. Adding to existing work on mass and spin measurements, two new variables for spin determination and an asymmetry for the determination of the couplings of the new particles are introduced. It is shown that these observables allow one to independently determine the spin and the couplings of the new particles, except for a few cases that turn out to be indistinguishable at the LHC. These findings are corroborated by results of an alternative analysis strategy based on an automated likelihood test.Comment: 18 pages, 3 figures, LaTe

Hadronic Cross-sections in two photon Processes at a Future Linear Collider

In this note we address the issue of measurability of the hadronic cross-sections at a future photon collider as well as for the two-photon processes at a future high energy linear $e^+e^-$ collider. We extend, to higher energy, our previous estimates of the accuracy with which the \gamgam\ cross-section needs to be measured, in order to distinguish between different theoretical models of energy dependence of the total cross-sections. We show that the necessary precision to discriminate among these models is indeed possible at future linear colliders in the Photon Collider option. Further we note that even in the $e^+e^-$ option a measurement of the hadron production cross-section via \gamgam processes, with an accuracy necessary to allow discrimination between different theoretical models, should be possible. We also comment briefly on the implications of these predictions for hadronic backgrounds at the future TeV energy $e^+e^-$ collider CLIC.Comment: 20 pages, 5 figures, LaTeX. Added an acknowledgemen