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Chloroplast function depends on the translocation of cytosolically synthesized precursor proteins into the organelle. The recognition and transfer of most precursor proteins across the outer membrane depend on a membrane inserted complex. Two receptor components of this complex, Toc34 and Toc159, are GTPases, which can be phosphorylated by kinases present in the hosting membrane. However, the physiological function of phosphorylation is not yet understood in detail. It is demonstrated that both receptors are phosphorylated within their G-domains. In vitro, the phosphorylation of Toc34 disrupts both homo- and heterodimerization of the G-domains as determined using a phospho-mimicking mutant. In endogenous membranes this mutation or phosphorylation of the wild-type receptor disturbs the association of Toc34, but not of Toc159 with the translocation pore. Therefore, phosphorylation serves as an inhibitor for the association of Toc34 with other components of the complex and phosphorylation can now be discussed as a mechanism to exchange different isoforms of Toc34 within this ensemble.
Gene trapping is used to introduce insertional mutations into genes of mouse embryonic stem cells (ESCs). It is performed with gene trap vectors that simultaneously mutate and report the expression of the endogenous gene at the site of insertion and provide a DNA tag for rapid identification of the disrupted gene. Gene traps have been employed worldwide to assemble libraries of mouse ESC lines harboring mutations in single genes, which can be used to make mutant mice. However, most of the employed gene trap vectors require gene expression for reporting a gene trap event and therefore genes that are poorly expressed may be under-represented in the existing libraries. To address this problem, we have developed a novel class of gene trap vectors that can induce gene expression at insertion sites, thereby bypassing the problem of intrinsic poor expression. We show here that the insertion of the osteopontin enhancer into several conventional gene trap vectors significantly increases the gene trapping efficiency in high-throughput screens and facilitates the recovery of poorly expressed genes.
Comparative studies suggest that at least some bird species have evolved mental skills similar to those found in humans and apes. This is indicated by feats such as tool use, episodic-like memory, and the ability to use one´s own experience in predicting the behavior of conspecifics. It is, however, not yet clear whether these skills are accompanied by an understanding of the self. In apes, self-directed behavior in response to a mirror has been taken as evidence of self-recognition. We investigated mirror-induced behavior in the magpie, a songbird species from the crow family. As in apes, some individuals behaved in front of the mirror as if they were testing behavioral contingencies. When provided with a mark, magpies showed spontaneous mark-directed behavior. Our findings provide the first evidence of mirror self-recognition in a non-mammalian species. They suggest that essential components of human self-recognition have evolved independently in different vertebrate classes with a separate evolutionary history.
Ribosome biogenesis in eukaryotes requires the participation of a large number of ribosome assembly factors. The highly conserved eukaryotic nucleolar protein Nep1 has an essential but unknown function in 18S rRNA processing and ribosome biogenesis. In Saccharomyces cerevisiae the malfunction of a temperature-sensitive Nep1 protein (nep1-1ts) was suppressed by the addition of S-adenosylmethionine (SAM). This suggests the participation of Nep1 in a methyltransferase reaction during ribosome biogenesis. In addition, yeast Nep1 binds to a 6-nt RNA-binding motif also found in 18S rRNA and facilitates the incorporation of ribosomal protein Rps19 during the formation of pre-ribosomes. Here, we present the X-ray structure of the Nep1 homolog from the archaebacterium Methanocaldococcus jannaschii in its free form (2.2 Å resolution) and bound to the S-adenosylmethionine analog S-adenosylhomocysteine (SAH, 2.15 Å resolution) and the antibiotic and general methyltransferase inhibitor sinefungin (2.25 Å resolution). The structure reveals a fold which is very similar to the conserved core fold of the SPOUT-class methyltransferases but contains a novel extension of this common core fold. SAH and sinefungin bind to Nep1 at a preformed binding site that is topologically equivalent to the cofactor-binding site in other SPOUT-class methyltransferases. Therefore, our structures together with previous genetic data suggest that Nep1 is a genuine rRNA methyltransferase.
Bency Eichorn learns in kollel and, on the side, has been researching about various segulos. For his wedding he authored a book, Simchas Zion, discussing the segulah of keeping the afikomom from year-to-year. The post below is a small part of a much larger project on this segulah and has been adapted for the blog.
Market uptake of pegylated interferons for the treatment of hepatitis C in Europe : meeting abstract
(2008)
Introduction and Objectives Hepatitis C virus (HCV) infection is a leading cause of chronic liver disease with life threatening sequelae such as end-stage liver cirrhosis and liver cancer. It is estimated that the infection annually causes about 86,000 deaths, 1.2 million disability adjusted life years (DALYs), and ¼ of the liver transplants in the WHO European region. Presently, only antiviral drugs can prevent the progression to severe liver disease. Pegylated interferons combined with ribavirin are considered as current state-of-the-art treatment. Objective of this investigation was to assess the market uptake of these drugs across Europe in order to find out whether there is unequal access to optimised therapy. Material and Methods We used IMS launch and sales data (April 2000 to December 2005) for peginterferons and ribavirin for 21 countries of the WHO European region. Market uptake was investigated by comparing the development of country-specific sales rates. For market access analysis, we converted sales figures into numbers of treated patients and related those to country-specific hepatitis C prevalence. To convert sales figures into patient figures, the amount of active pharmaceutical ingredients (API) sold was divided by average total patient doses (ATPD), derived by a probability tree-based calculation algorithm accounting for genotype distribution, early stopping rules, body weight, unscheduled treatment stops and dose reductions Ntotal=APIPegIFNalpha-2a/ATPDPegIFNalpha-2a+APIPegIFN&alpha-2b/ATPDPegIFNalpha-2b For more concise result presentation the 21 included countries were aggregated into four categories: 1. EU founding members (1957): Belgium, France, Germany, Italy and Netherlands; 2. Countries joining EU before 2000: Austria (1995), Denmark (1973), Finland (1995), Greece (1981), Republic of Ireland (1973), Spain (1986), Sweden and UK (1973) 3. Countries joining EU after 2000: Czech Republic (2004), Hungary (2004), Poland (2004) and Romania (2007); 4. EU non-member states: Norway, Russia, Switzerland and Turkey. Results Market launch and market uptake of the investigated drugs differed considerably across countries. The earliest, most rapid and highest increases in sales rates were observed in the EU founding member states, followed by countries that joined the EU before 2000, countries that joined the EU after 2000, and EU non-member states. Most new EU member states showed a noticeable increase in sales after joining the EU. Market access analysis yielded that until end of 2005, about 308 000 patients were treated with peginterferon in the 21 countries. Treatment rates differed across Europe. The number of patients ever treated with peginterferon per 100 prevalent cases ranged from 16 in France to less than one in Romania, Poland, Greece and Russia. Discussion Peginterferon market uptake and prevalence adjusted treatment rates were found to vary considerably across 21 countries in the WHO European region suggesting unequal access to optimised therapy. Poor market access was especially common in low-resource countries. Besides budget restrictions, national surveillance and prevention policy should be considered as explanations for market access variation. Although our results allowed for the ranking of countries in order of market access, no final conclusions on over- or undertreatment can be drawn, because the number of patients who really require antiviral treatment is unknown. Further research based on pan-European decision models is recommended to determine the fraction of not yet successfully treated but treatable patients among those ever diagnosed with HCV. ...
Many new gene copies emerged by gene duplication in hominoids, but little is known with respect to their functional evolution. Glutamate dehydrogenase (GLUD) is an enzyme central to the glutamate and energy metabolism of the cell. In addition to the single, GLUD-encoding gene present in all mammals (GLUD1), humans and apes acquired a second GLUD gene (GLUD2) through retroduplication of GLUD1, which codes for an enzyme with unique, potentially brain-adapted properties. Here we show that whereas the GLUD1 parental protein localizes to mitochondria and the cytoplasm, GLUD2 is specifically targeted to mitochondria. Using evolutionary analysis and resurrected ancestral protein variants, we demonstrate that the enhanced mitochondrial targeting specificity of GLUD2 is due to a single positively selected glutamic acid-to-lysine substitution, which was fixed in the N-terminal mitochondrial targeting sequence (MTS) of GLUD2 soon after the duplication event in the hominoid ancestor ~18–25 million years ago. This MTS substitution arose in parallel with two crucial adaptive amino acid changes in the enzyme and likely contributed to the functional adaptation of GLUD2 to the glutamate metabolism of the hominoid brain and other tissues. We suggest that rapid, selectively driven subcellular adaptation, as exemplified by GLUD2, represents a common route underlying the emergence of new gene functions.
C2-symmetric bisamidines : chiral Brønsted bases catalysing the Diels-Alder reaction of anthrones
(2008)
C2-symmetric bisamidines 8 have been tested as chiral Brønsted bases in the Diels- Alder reaction of anthrones and N-substituted maleimides. High yields of cycloadducts and significant asymmetric inductions up to 76% ee are accessible. The proposed mechanism involves proton transfer between anthrone and bisamidine, association of the resulting ions and finally a cycloaddition step stereoselectively controlled by the chiral ion pair.
Oscillatory activity in human electro- or magnetoencephalogram has been related to cortical stimulus representations and their modulation by cognitive processes. Whereas previous work has focused on gamma-band activity (GBA) during attention or maintenance of representations, there is little evidence for GBA reflecting individual stimulus representations. The present study aimed at identifying stimulus-specific GBA components during auditory spatial short-term memory. A total of 28 adults were assigned to 1 of 2 groups who were presented with only right- or left-lateralized sounds, respectively. In each group, 2 sample stimuli were used which differed in their lateralization angles (15° or 45°) with respect to the midsagittal plane. Statistical probability mapping served to identify spectral amplitude differences between 15° versus 45° stimuli. Distinct GBA components were found for each sample stimulus in different sensors over parieto-occipital cortex contralateral to the side of stimulation peaking during the middle 200–300 ms of the delay phase. The differentiation between "preferred" and "nonpreferred" stimuli during the final 100 ms of the delay phase correlated with task performance. These findings suggest that the observed GBA components reflect the activity of distinct networks tuned to spatial sound features which contribute to the maintenance of task-relevant information in short-term memory.
Cytotoxic T-lymphocytes play an important role in the protection against viral infections, which they detect through the recognition of virus-derived peptides, presented in the context of MHC class I molecules at the surface of the infected cell. The transporter associated with antigen processing (TAP) plays an essential role in MHC class I–restricted antigen presentation, as TAP imports peptides into the ER, where peptide loading of MHC class I molecules takes place. In this study, the UL49.5 proteins of the varicelloviruses bovine herpesvirus 1 (BHV-1), pseudorabies virus (PRV), and equine herpesvirus 1 and 4 (EHV-1 and EHV-4) are characterized as members of a novel class of viral immune evasion proteins. These UL49.5 proteins interfere with MHC class I antigen presentation by blocking the supply of antigenic peptides through inhibition of TAP. BHV-1, PRV, and EHV-1 recombinant viruses lacking UL49.5 no longer interfere with peptide transport. Combined with the observation that the individually expressed UL49.5 proteins block TAP as well, these data indicate that UL49.5 is the viral factor that is both necessary and sufficient to abolish TAP function during productive infection by these viruses. The mechanisms through which the UL49.5 proteins of BHV-1, PRV, EHV-1, and EHV-4 block TAP exhibit surprising diversity. BHV-1 UL49.5 targets TAP for proteasomal degradation, whereas EHV-1 and EHV-4 UL49.5 interfere with the binding of ATP to TAP. In contrast, TAP stability and ATP recruitment are not affected by PRV UL49.5, although it has the capacity to arrest the peptide transporter in a translocation-incompetent state, a property shared with the BHV-1 and EHV-1 UL49.5. Taken together, these results classify the UL49.5 gene products of BHV-1, PRV, EHV-1, and EHV-4 as members of a novel family of viral immune evasion proteins, inhibiting TAP through a variety of mechanisms.
The degradation of the poly(A) tail is crucial for posttranscriptional gene regulation and for quality control of mRNA. Poly(A)-specific ribonuclease (PARN) is one of the major mammalian 3’ specific exo-ribonucleases involved in the degradation of the mRNA poly(A) tail, and it is also involved in the regulation of translation in early embryonic development. The interaction between PARN and the m7GpppG cap of mRNA plays a key role in stimulating the rate of deadenylation. Here we report the solution structures of the cap-binding domain of mouse PARN with and without the m7GpppG cap analog. The structure of the cap-binding domain adopts the RNA recognition motif (RRM) with a characteristic a-helical extension at its C-terminus, which covers the b-sheet surface (hereafter referred to as PARN RRM). In the complex structure of PARN RRM with the cap analog, the base of the N7-methyl guanosine (m7G) of the cap analog stacks with the solvent-exposed aromatic side chain of the distinctive tryptophan residue 468, located at the C-terminal end of the second b-strand. These unique structural features in PARN RRM reveal a novel cap-binding mode, which is distinct from the nucleotide recognition mode of the canonical RRM domains.
We performed a bioinformatical analysis of protein export elements (PEXEL) in the putative proteome of the malaria parasite Plasmodium falciparum. A protein family-specific conservation of physicochemical residue profiles was found for PEXEL-flanking sequence regions. We demonstrate that the family members can be clustered based on the flanking regions only and display characteristic hydrophobicity patterns. This raises the possibility that the flanking regions may contain additional information for a family-specific role of PEXEL. We further show that signal peptide cleavage results in a positional alignment of PEXEL from both proteins with, and without, a signal peptide.
While the adaptor SKAP-55 mediates LFA-1 adhesion on T-cells, it is not known whether the adaptor regulates other aspects of signaling. SKAP-55 could potentially act as a node to coordinate the modulation of adhesion with downstream signaling. In this regard, the GTPase p21ras and the extracellular signal-regulated kinase (ERK) pathway play central roles in T-cell function. In this study, we report that SKAP-55 has opposing effects on adhesion and the activation of the p21ras -ERK pathway in T-cells. SKAP-55 deficient primary T-cells showed a defect in LFA-1 adhesion concurrent with the hyper-activation of the ERK pathway relative to wild-type cells. RNAi knock down (KD) of SKAP-55 in T-cell lines also showed an increase in p21ras activation, while over-expression of SKAP-55 inhibited activation of ERK and its transcriptional target ELK. Three observations implicated the p21ras activating exchange factor RasGRP1 in the process. Firstly, SKAP-55 bound to RasGRP1 via its C-terminus, while secondly, the loss of binding abrogated SKAP-55 inhibition of ERK and ELK activation. Thirdly, SKAP-55−/− primary T-cells showed an increased presence of RasGRP1 in the trans-Golgi network (TGN) following TCR activation, the site where p21ras becomes activated. Our findings indicate that SKAP-55 has a dual role in regulating p21ras-ERK pathway via RasGRP1, as a possible mechanism to restrict activation during T-cell adhesion.
Diabrotica virgifera virgifera LeConte, in its original North American habitat also known as western corn rootworm beetle, actively continues its expansion to new territories and uses Homo sapiens as its prime vector. It took only 15 years to spread to and occupy the southeastern and central parts of Europe, so far with the exception of Denmark where it has not been documented as of 2007. Economic thresholds have been reached and surpassed only in Southeast European countries like Slovakia, Hungary, Serbia, Eastern Croatia, Romania and Northern Italy. But both, the area affected and the severity of symptoms are increasing. Model calculations by a number of authors (Baufeld & Enzian, 2005 a and b; Hongmei Li & al. 2006, CLIMEX model) indicate a definitive propensity of D. v. virgifera to expand its currently occupied territory to regions with moderate temperatures and Zea mays cultivation. East Africa and Eastern Asia are included in the list of potential candidates for future inadvertent introduction. In most discussions it is tacitly and erroneously assumed that Z. mays is the only or the only important host of D. v. virgifera. Our recent observations in Eastern Slovenia on the oil pumpkin Cucurbita pepo indicate, however, that this simplifying assumption is notlonger strictly valid. It has to be modified in light of new evidence. Here, we report a few field experiments conducted in August of 2006 clarifying the host status of C. pepo in a European country.
Der inzwischen auf der EPPO A1-Liste aufgeführte Quarantäneschädling, der Bockkäfer Anoplophora glabripennis (Motschulsky, 1853), ist seit seiner Einschleppung nach Braunau am Inn (Österreich) im Jahr 2001 in ganz Europa ein Begriff. Neben zwei unabhängigen Fundorten in Frankreich, 2003 in Gien und 2004 in Sainte-Anne-sur-Brivet (Hérard & al. 2005) wurde eine Freiland-Population dieser Laubbäume schädigenden Art erstmals 2004 auch in Deutschland festgestellt. Bisherige Funde von Käfern in Deutschland waren auf das Gelände des Hamburger Hafens, Umschlagplatz vieler Container aus dem asiatischen Raum, beschränkt (Schliesske 2001). Es konnte aber nie eine Übersiedelung auf Laubbäume in der Umgebung beobachtet werden. Die Situation änderte sich gravierend, als offensichtlich befallene Bäume in Neukirchen am Inn (Gemeinde Neuburg am Inn, Landkreis Passau) in Bayern festgestellt wurden. Symptome wie mangelnde Belaubung, kraterförmige Eiablagestellen und Austritt von Bohrmehl an einem Ahornbaum deuteten auf Befall mit A. glabripennis hin. Nach Entnahme von Astteilen wurden in Gängen Bockkäfer-Larven gefunden, die von Kollegen des Bundesamtes und Forschungszentrums für Wald (BFW) in Wien mittels DNA-Analyse zweifelsfrei als A. glabripennis identifiziert wurden (Hoyer & al. 2003).
The codling moth, Cydia pomonella (Lep., Tortricidae), is a significant pest of orchard crops such as apple and pear in Southern Germany, and can cause severe economic damage to apple crops. Due to resistance to conventional pesticides and the growing market for organic fruit, Cydia pomonella Granulovirus (CpGV) has been used to control C. pomonella in Germany for over 10 years. Recently, populations exhibiting resistance to CpGV have been reported. In this study, we have used amplified fragment length polymorphism (AFLP) markers to estimate genetic variations between eight different C. pomonella populations, which were obtained from different locations exhibiting varying levels of resistance to CpGV. Three different AFLP primer combinations generated a total of 194 AFLP fragments, ranging from 57.84 to 424.11 bp, with an average of 59.23 amplified fragments per primer combination. The total number of segregating fragments ranged from 181 to 115 and resulted in a high loci polymorphism of 100% in most cases, except for two populations, where it was found to be 88.1% and 93.3%. An analysis of genetic variation based on the obtained AFLP markers resulted in high gene diversity (Hj) values, ranging between 0.2884 to 0.3508. Hj values also indicated a loss in gene diversity within a population over time. The Wright Fixation Index (FST) values indicated a low to moderate genetic differentiation in the populations. The cluster analysis (UPGMA), based on genetic distance values, showed that the majority of C. pomonella populations from different locations were clearly distributed into distinct groups and showed a large genetic variability.
Neozoa werden als gebietsfremde Arten definiert, welche nach 1492 direkt oder indirekt durch den Menschen eingeführt wurden und selbstreproduzierende Populationen zu bilden im Stande sind (Essl & Rabitsch 2002). Biologische Invasionen sind schon seit dem Zeitalter des Kolonialismus ein bekanntes Phänomen, gegenwärtig hat das Problem in einer Zeit der Globalisierung von Wirtschaft und Tourismus und einer globalen Klimaveränderung jedoch eine neue Dimension und Geschwindigkeit erreicht. In Österreich gibt es grundsätzlich eine gute Datenbasis über das Auftreten von benthischen Einwanderern. Bereits in der älteren Literatur findet man dazu einige Hinweise, z.B.: Strouhal (1939), Vornatscher (1965) und Liepolt (1965–67). Größere Aufmerksamkeit wurde dem Phänomen invasiver Arten seit der massiven Ausbreitung der Dreikantmuschel Dreissena polymorpha geschenkt, als sich diese in den 1970er Jahren in vielen Flüssen und Seen in einem sehr kurzen Zeitraum ausbreitete. Verstärkte Beachtung wurde den Neozoa in der Limnologie aber erst im letzten Jahrzehnt zuteil, als die Fundnachweise immer häufiger wurden und sowohl ökonomische als auchbemerkenswerte ökologische Auswirkungen durch invasive Arten vermutet oder nachgewiesen wurden. Beispielsweise belegte Gruber (2006), dass der autochthone Steinkrebs (Austropotamobius torrentium) bereits bis in die Quellregion hinein durch den amerikanischen Signalkrebs (Pacifastacus leniusculus) verdrängt wird. In Österreich entstand als Folge des erhöhten Problembewusstseins in den letzten Jahren eine Reihe von umfassenden Publikationen zum Thema Neobiota: „Invaders“ (Aescht & al. 1995), „Neobiota in Österreich“ (Essl & Rabitsch 2002) und „Aliens“ (Wallner 2005). Während die überwiegende Zahl der Publikationen zum Thema Neozoa von taxonomischen Fachspezialisten stammt und deshalb zumeist auf eine oder einige ausgewählte Tiergruppen beschränkt bleibt, basiert vorliegender Beitrag auf Monitoringdaten und gewässerökologischen Untersuchungen, in welchen immer die gesamte Artengesellschaft der benthischen Evertebraten erhoben wurde.
Die 31. Karl-Medaille wurde vor 2 Jahren Herrn Professor Francke, Hamburg verliehen; es ist vielleicht kein Zufall, dass wir heute das Ehepaar Levinson ehren dürfen. verbindet sie doch eine langjährige und vor allen Dingen sehr erfolgreiche Zusammenarbeit. Hierzu ein Beispiel: Das Weibchen des Tabakkäfers Lasioderma serricorne produziert das Sexualpheromon Serricornin. Gemeinsam mit Herrn Francke ist Ihnen die Aufklärung der seltenen Speicherform eines Pheromons, und zwar des Anhydroserricornins, gelungen. Sie haben die Pheromondrüse beschrieben und die biologische Aktivität der Enantiomeren ermittelt. Basierend auf diesen Erkenntnissen entwickelten Sie gemeinsam mit British American Tobacco eine Lockstoff-Falle, die von Ihnen unter Praxisbedingungen in Tabaklagern in Deutschland, Griechenland, Zypern und Ägypten getestet wurde. Ergebnis ist das Produkt Lasiotrap, das kommerziell zu Monitoring und Massenfang des Tabakkäfers eingesetzt wird. Mit diesem Beispiel möchte ich einen wesentlichen Ansatz Ihres wissenschaftlichen Denkens und Forschens beschreiben. ...
Sie sehen und hören recht – die im Foto gezeigte, heute zu ehrende Dame begeht in einigen Monaten ihren 85. Geburtstag und startet in wenigen Tagen zu ihrer nächsten entomologischen Sammelreise nach Namibia. Sie entschuldigt sich deshalb, dass sie bei der Verleihung der bedeutendsten deutschen Auszeichnung für Leistungen entomologischer Forschung, der Fabricius-Medaille, für ihr bisheriges Lebenswerk, heute nicht anwesend sein kann. Ich hätte ihr gesagt: Liebe Ursula, ich kannte Deinen Namen schon aus der Fachliteratur, bevor ich Dich das erste Mal Mitte der achtziger Jahre bei einer der jährlichen Tagungen der Arbeitsgemeinschaft der Mitteleuropäischen Heteropterologen im „Westen“ persönlich traf und überrascht war, einer agilen rüstigen Seniorin zu begegnen. Seither sind wir mehrmals zusammengetroffen und ich erinnere...