Slower rescue of ER homeostasis by the unfolded protein response pathway associated with common variable immunodeficiency

Common variable immunodeficiency (CVID) is a primary immunodeficiency characterized by hypogammaglobulinemia and recurrent infections. Herein we addressed the role of unfolded protein response (UPR) in the pathogenesis of the disease. Augmented unspliced X-box binding protein 1 (XBP-1) mRNA concurrent with co-localization of IgM and BiP/GRP78 were found in one CVID patient. At confocal microscopy analysis this patient`s cells were enlarged and failed to present the typical surface distribution of IgM, which accumulated within an abnormally expanded endoplasmic reticulum. Sequencing did not reveal any mutation on XBP-1, neither on IRE-1 alpha that could potentially prevent the splicing to occur. Analysis of spliced XBP-1, IRE-1 alpha and BiP messages after LPS or Brefeldin A treatment showed that, unlike healthy controls that respond to these endoplasmic reticulum (ER) stressors by presenting waves of transcription of these three genes, this patient`s cells presented lower rates of transcription, not reaching the same level of response of healthy subjects even after 48 h of ER stress. Treatment with DMSO rescued IgM and IgG secretion as well as the expression of spliced XBP-1. Our findings associate diminished splicing of XBP-1 mRNA with accumulation of IgM within the ER and lower rates of chaperone transcription, therefore providing a mechanism to explain the observed hypogammaglobulinemia. (C) 2008 Elsevier Ltd. All rights reserved.

Alternative diagnostic diagrams and the `forgotten` population of weak line galaxies in the SDSS

A numerous population of weak line galaxies (WLGs) is often left out of statistical studies on emission-line galaxies (ELGs) due to the absence of an adequate classification scheme, since classical diagnostic diagrams, such as [O iii]/H beta versus [N ii]/H alpha (the BPT diagram), require the measurement of at least four emission lines. This paper aims to remedy this situation by transposing the usual divisory lines between star-forming (SF) galaxies and active galactic nuclei (AGN) hosts and between Seyferts and LINERs to diagrams that are more economical in terms of line quality requirements. By doing this, we rescue from the classification limbo a substantial number of sources and modify the global census of ELGs. More specifically, (1) we use the Sloan Digital Sky Survey Data Release 7 to constitute a suitable sample of 280 000 ELGs, one-third of which are WLGs. (2) Galaxies with strong emission lines are classified using the widely applied criteria of Kewley et al., Kauffmann et al. and Stasinska et al. to distinguish SF galaxies and AGN hosts and Kewley et al. to distinguish Seyferts from LINERs. (3) We transpose these classification schemes to alternative diagrams keeping [N ii]/H alpha as a horizontal axis, but replacing H beta by a stronger line (H alpha or [O ii]), or substituting the ionization-level sensitive [O iii]/H beta ratio with the equivalent width of H alpha (W(H alpha)). Optimized equations for the transposed divisory lines are provided. (4) We show that nothing significant is lost in the translation, but that the new diagrams allow one to classify up to 50 per cent more ELGs. (5) Introducing WLGs in the census of galaxies in the local Universe increases the proportion of metal-rich SF galaxies and especially LINERs. In the course of this analysis, we were led to make the following points. (i) The Kewley et al. BPT line for galaxy classification is generally ill-used. (ii) Replacing [O iii]/H beta by W(H alpha) in the classification introduces a change in the philosophy of the distinction between LINERs and Seyferts, but not in its results. Because the W(H alpha) versus [N ii]/H alpha diagram can be applied to the largest sample of ELGs without loss of discriminating power between Seyferts and LINERs, we recommend its use in further studies. (iii) The dichotomy between Seyferts and LINERs is washed out by WLGs in the BPT plane, but it subsists in other diagnostic diagrams. This suggests that the right wing in the BPT diagram is indeed populated by at least two classes, tentatively identified with bona fide AGN and `retired` galaxies that have stopped forming stars and are ionized by their old stellar populations.

Over-expression of COQ10 in Saccharomyces cerevisiae inhibits mitochondrial respiration

COQ10 deletion in Saccharomyces cerevisiae elicits a defect in mitochondrial respiration correctable by addition of coenzyme Q(2). Rescue of respiration by Q(2) is a characteristic of mutants blocked in coenzyme Q(6) synthesis. Unlike Q(6) deficient mutants, mitochondria of the coq10 null mutant have wild-type concentrations Of Q(6). The physiological significance of earlier observations that purified Coq10p contains bound Q(6) was examined in the present study by testing the in vivo effect of over-expression of Coq10p on respiration. Mitochondria with elevated levels of Coq10p display reduced respiration in the bc1 span of the electron transport chain, which can be restored with exogenous Q(2). This suggests that in vivo binding of Q(6) by excess Coq10p reduces the pool of this redox carrier available for its normal function in providing electrons to the bc1 complex. This is confirmed by observing that extra Coq8p relieves the inhibitory effect of excess Coq10p. Coq8p is a putative kinase, and a high-copy suppressor of the coq10 null mutant. As shown here, when over-produced in coq mutants, Coq8p counteracts turnover of Coq3p and Coq4p subunits of the Q-biosynthetic complex. This can account for the observed rescue by COQ8 of the respiratory defect in strains over-producing Coq10p. (C) 2010 Elsevier Inc. All rights reserved.

The catalytic and other residues essential for the activity of the midgut trehalase from Spodoptera frugiperda

Trehalase (EC 3.2.1.28) hydrolyzes only alpha, alpha`- trehalose and is present in a variety of organisms, but is most important in insects and fungi. Crystallographic data showed that bacterial trehalase has 0312 and E496 as the catalytical residues and three Arg residues in the active site. Those residues have homologous in all family 37 trehalases including Spodoptera frugiperda trehalase (0322, E520, R169, R227, R287). To test the role of these residues, mutants of trehalase were produced. All mutants were at least four orders of magnitude less active than wild type trehalase and no structural difference between these mutants and wild type enzyme were discernible by circular dichroism. D322A and E520 pH-activity profile lacked the alkaline arm and the acid arm, respectively, suggesting that D322 is the acid and E520 the basic catalyst. Azide increases E520A activity three times, confirming its action as the basic catalyst. Taking into account the decrease in activity after substitution for alanine residue, the three arginine residues are as important as the catalytical ones to trehalase activity. This clarifies the previous misidentification of an Arg residue as the acid catalyst. As far as we know, this is the first report on the functional identification residues important for trehalase activity. (C) 2010 Elsevier Ltd. All rights reserved.