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11.
TRAPP, a highly conserved novel complex on the cis-Golgi that mediates vesicle docking and fusion. 总被引:16,自引:0,他引:16 下载免费PDF全文
M Sacher Y Jiang J Barrowman A Scarpa J Burston L Zhang D Schieltz J R Yates rd H Abeliovich S Ferro-Novick 《The EMBO journal》1998,17(9):2494-2503
We previously identified BET3 by its genetic interactions with BET1, a gene whose SNARE-like product acts in endoplasmic reticulum (ER)-to-Golgi transport. To gain insight into the function of Bet3p, we added three c-myc tags to its C-terminus and immunopurified this protein from a clarified detergent extract. Here we report that Bet3p is a member of a large complex ( approximately 800 kDa) that we call TRAPP (transport protein particle). We propose that TRAPP plays a key role in the targeting and/or fusion of ER-to-Golgi transport vesicles with their acceptor compartment. The localization of Bet3p to the cis-Golgi complex, as well as biochemical studies showing that Bet3p functions on this compartment, support this hypothesis. TRAPP contains at least nine other constituents, five of which have been identified and shown to be highly conserved novel proteins. 相似文献
12.
Sacher F Jais P Stephenson K O'Neill MD Hocini M Clementy J Stevenson WG Haissaguerre M 《Indian pacing and electrophysiology journal》2007,7(1):1-6
Phrenic Nerve Injury (PNI) has been well studied by cardiac surgeons. More recently it has been recognized as a potential complication of catheter ablation with a prevalence of 0.11 to 0.48 % after atrial fibrillation (AF) ablation. This review will focus on PNI after AF ablation. Anatomical studies have shown a close relationship between the right phrenic nerve and it's proximity to the superior vena cava (SVC), and the antero-inferior part of the right superior pulmonary vein (RSPV). In addition, the proximity of the left phrenic nerve to the left atrial appendage has been well established. Independent of the type of ablation catheter (4 mm, 8 mm, irrigated tip, balloon) or energy source used (radiofrequency (RF), ultrasound, cryothermia, and laser); the risk of PNI exists during ablation at the critical areas listed above. Although up to thirty-one percent of patients with PNI after AF ablation remain asymptomatic, dyspnea remain the cardinal symptom and is present in all symptomatic patients. Despite the theoretical risk for significant adverse effect on functional status and quality of life, short-term outcomes from published studies appear favorable with 81% of patients with PNI having a complete recovery after 7 +/- 7 months. CONCLUSION: Existing studies have described PNI as an uncommon but avoidable complication in patients undergoing pulmonary vein isolation for AF. Prior to ablation at the SVC, antero-inferior RSPV ostium or the left atrial appendage, pacing should be performed before energy delivery. If phrenic nerve capture is documented, energy delivery should be avoided at this site. Electrophysiologist's vigilance as well as pacing prior to ablation at high risk sites in close proximity to the phrenic nerve are the currently available tools to avoid the complication of PNI. 相似文献
13.
Hasib L Dilcher M Hufert F Meyer-König U König-Meyer U Weidmann M 《Molecular biotechnology》2011,49(2):176-186
It is suspected that apart from tick-borne encephalitis virus several additional European Arboviruses such as the sandfly
borne Toscana virus, sandfly fever Sicilian virus and sandfly fever Naples virus, mosquito-borne Tahyna virus, Inkoo virus,
Batai virus and tick-borne Uukuniemi virus cause aseptic meningo-encephalitis or febrile disease in Europe. Currently, the
microarray technology is developing rapidly and there are many efforts to apply it to infectious diseases diagnostics. In
order to arrive at an assay system useful for high throughput analysis of samples from aseptic meningo-encephalitis cases
the authors developed a combined multiplex ligation-dependent probe amplification and flow-through microarray assay for the
detection of European Bunyaviruses. These results show that this combined assay indeed is highly sensitive, and specific for
the accurate detection of multiple viruses. 相似文献
14.
RNAi screen of Salmonella invasion shows role of COPI in membrane targeting of cholesterol and Cdc42
Raphael Sacher Berend Snijder Markus Schlumberger Samuel Rout Manuel Stark Christian von Mering Lucas Pelkmans Wolf‐Dietrich Hardt 《Molecular systems biology》2011,7(1)
The pathogen Salmonella Typhimurium is a common cause of diarrhea and invades the gut tissue by injecting a cocktail of virulence factors into epithelial cells, triggering actin rearrangements, membrane ruffling and pathogen entry. One of these factors is SopE, a G‐nucleotide exchange factor for the host cellular Rho GTPases Rac1 and Cdc42. How SopE mediates cellular invasion is incompletely understood. Using genome‐scale RNAi screening we identified 72 known and novel host cell proteins affecting SopE‐mediated entry. Follow‐up assays assigned these ‘hits’ to particular steps of the invasion process; i.e., binding, effector injection, membrane ruffling, membrane closure and maturation of the Salmonella‐containing vacuole. Depletion of the COPI complex revealed a unique effect on virulence factor injection and membrane ruffling. Both effects are attributable to mislocalization of cholesterol, sphingolipids, Rac1 and Cdc42 away from the plasma membrane into a large intracellular compartment. Equivalent results were obtained with the vesicular stomatitis virus. Therefore, COPI‐facilitated maintenance of lipids may represent a novel, unifying mechanism essential for a wide range of pathogens, offering opportunities for designing new drugs. 相似文献
15.
Kim MS Yi MJ Lee KH Wagner J Munger C Kim YG Whiteway M Cygler M Oh BH Sacher M 《Traffic (Copenhagen, Denmark)》2005,6(12):1183-1195
Transport protein particle (TRAPP) comprises a family of two highly related multiprotein complexes, with seven common subunits, that serve to target different classes of transport vesicles to their appropriate compartments. Defining the architecture of the complexes will advance our understanding of the functional differences between these highly related molecular machines. Genetic analyses in yeast suggested a specific interaction between the TRAPP subunits Bet3p and Trs33p. A mammalian bet3-trs33 complex was crystallized, and the structure was solved to 2.2 angstroms resolution. Intriguingly, the overall fold of the bet3 and trs33 monomers was similar, although the proteins had little overall sequence identity. In vitro experiments using yeast TRAPP subunits indicated that Bet3p binding to Trs33p facilitates the interaction between Bet3p and another TRAPP subunit, Bet5p. Mutational analysis suggests that yeast Trs33p facilitates other Bet3p protein-protein interactions. Furthermore, we show that Trs33p can increase the Golgi-localized pool of a mutated Bet3 protein normally found in the cytosol. We propose that one of the roles of Trs33p is to facilitate the incorporation of the Bet3p subunit into assembling TRAPP complexes. 相似文献
16.
17.
The transport of material between membrane-bounded organelles in eukaryotic cells requires the accurate delivery of different classes of carrier vesicles to specific target compartments. Recent studies indicate that different targeting reactions involve distinct protein complexes that act to mark the target organelle for incoming vesicles. This review focuses on the proteins and protein complexes that have been implicated in various targeting reactions. 相似文献
18.
S Vargas A Schuster K Sacher G Büttner S Schätzle B Läuchli K Hall JN Hooper D Erpenbeck G Wörheide 《PloS one》2012,7(7):e39345
Background
Phylum Porifera includes ∼8,500 valid species distributed world-wide in aquatic ecosystems ranging from ephemeral fresh-water bodies to coastal environments and the deep-sea. The taxonomy and systematics of sponges is complicated, and morphological identification can be both time consuming and erroneous due to phenotypic convergence and secondary losses, etc. DNA barcoding can provide sponge biologists with a simple and rapid method for the identification of samples of unknown taxonomic membership. The Sponge Barcoding Project (www.spongebarcoding.org), the first initiative to barcode a non-bilaterian metazoan phylum, aims to provide a comprehensive DNA barcode database for Phylum Porifera.Methodology/Principal Findings
∼7,400 sponge specimens have been extracted, and amplification of the standard COI barcoding fragment has been attempted for approximately 3,300 museum samples with ∼25% mean amplification success. Based on this comprehensive sampling, we present the first report on the workflow and progress of the sponge barcoding project, and discuss some common pitfalls inherent to the barcoding of sponges.Conclusion
A DNA-barcoding workflow capable of processing potentially large sponge collections has been developed and is routinely used for the Sponge Barcoding Project with success. Sponge specific problems such as the frequent co-amplification of non-target organisms have been detected and potential solutions are currently under development. The initial success of this innovative project have already demonstrated considerable refinement of sponge systematics, evaluating morphometric character importance, geographic phenotypic variability, and the utility of the standard barcoding fragment for Porifera (despite its conserved evolution within this basal metazoan phylum). 相似文献19.
Dirk Erpenbeck Kathryn Hall Belinda Alvarez Gabriele Büttner Katharina Sacher Simone Schätzle Astrid Schuster Sergio Vargas John N. A. Hooper Gert Wörheide 《Organisms Diversity & Evolution》2012,12(1):57-70
Halichondrid sponges play a pivotal role in the classification of demosponges as changes in their classification has had direct
consequences for the classification of Demospongiae. Historically, the systematics of halichondrids has been unstable. During
the 1950s, the order was divided into two subclasses, which were based on empirical and assumed reproductive data. Subsequent
morphological and biochemical analyses postulated the re-merging of halichondrid families, but recent molecular data indicate
their polyphyly. Here we review the classification history of halichondrid taxa, compare it with the current and predominantly
ribosomal molecular data, and support the new phylogenetic hypotheses with mitochondrial data from DNA barcoding. 相似文献
20.
A Wawrzeńczyk A Sacher M Mac M J Na?ecz K A Na?ecz 《European journal of biochemistry》2001,268(7):2091-2098
The accumulation of carnitine was measured in cerebral cortex neurons isolated from adult rat brain. This process was found to be lowered by 40% after preincubation with ouabain and with SH-group reagents (N-ethylmaleimide and mersalyl). The initial velocity of carnitine transport was found to be inhibited by 4-aminobutyrate (GABA) in a competitive way (Ki = 20.9 +/- 2.4 mM). However, of various inhibitors of GABA transporters, only nipecotic acid and very high concentrations of 1-[2-([(diphenylmethylene)amino]oxy)ethyl]-1,2,5,6-tetrahydro-3-pyridinecarboxylic acid hydrochloride (NO-711) acid decreased carnitine accumulation while betaine, taurine and beta-alanine had no effect. The GABA transporters expressed in Xenopus laevis oocytes did not transport carnitine. Moreover, carnitine was not observed to diminish the accumulation of GABA in cerebral cortex neurons, which further excluded a possible involvement of the GABA transporter GAT1 in the process of carnitine accumulation, despite the expression of this protein in the cells under study. The absence of carnitine transporter OCTN2 in rat cerebral cortex neurons (K. A. Na?ecz, D. Dymna, J. E. Mroczkowska, A. Bro?r, S. Bro?r, M. J. Na?ecz and R. Cecchelli, unpublished results), together with the insensitivity of carnitine accumulation towards betaines, implies that a novel transporting protein is present in these cells. 相似文献