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Ilmenauer Beiträge zur Wirtschaftsinformatik Nr. 2011-02 / Technische Universität Ilmenau, Fakultät für Wirtschaftswissenschaften, Institut für Wirtschaftsinformatik ISSN 1861-9223 ISBN 978-3-938940-33-

Functional characterization of VirB/VirD4 and Icm/Dot type IV secretion systems from the plant-pathogenic bacterium Xanthomonas euvesicatoria

IntroductionMany Gram-negative plant- and animal-pathogenic bacteria employ type IV secretion (T4S) systems to transport proteins or DNA/protein complexes into eukaryotic or bacterial target cells. T4S systems have been divided into minimized and expanded T4S systems and resemble the VirB/VirD4 T4S system from the plant pathogen Agrobacterium tumefaciens and the Icm/Dot T4S system from the human pathogen Legionella pneumophila, respectively. The only known plant pathogen with both types of T4S systems is Xanthomonas euvesicatoria which is the causal agent of bacterial spot disease on pepper and tomato plants.Results and discussionIn the present study, we show that virB/virD4 and icm/dot T4S genes are expressed and encode components of oligomeric complexes corresponding to known assemblies of VirB/VirD4 and Icm/Dot proteins. Both T4S systems are dispensable for the interaction of X. euvesicatoria with its host plants and do not seem to confer contact-dependent lysis of other bacteria, which was previously shown for the chromosomally encoded VirB/VirD4 T4S system from Xanthomonas axonopodis pv. citri. The corresponding chromosomal T4S gene cluster from X. euvesicatoria is incomplete, however, the second plasmid-localized vir gene cluster encodes a functional VirB/VirD4 T4S system which contributes to plasmid transfer. In agreement with this finding, we identified the predicted relaxase TraI as substrate of the T4S systems from X. euvesicatoria. TraI and additional candidate T4S substrates with homology to T4S effectors from X. axonopodis pv. citri interact with the T4S coupling protein VirD4. Interestingly, however, the predicted C-terminal VirD4-binding sites are not sufficient for T4S, suggesting the contribution of additional yet unknown mechanisms to the targeting of T4S substrates from X. euvesicatoria to both VirB/VirD4 and Icm/Dot T4S systems

Arbeitsbericht Nr. 2014-03, Juli 2014

Unternehmen wenden einen großen Teil des IT-Budgets für die Integration von Informationssystemen auf. Problematisch ist, dass die Zusammensetzung der Integrationskosten häufig nicht hinreichend bekannt ist und in der Literatur keine eindeutige Definition der Integrationskosten existiert, was die Analyse der Wirtschaftlichkeit von Integrationsprojekten erheblich erschwert. Um eine Spezifizierung des Begriffs der Integrationskosten vorzunehmen, werden in der Literatur diskutierte Konzepte analysiert und gegenübergestellt. Im Anschluss wird eine Kostenmatrix erstellt, mit deren Hilfe Unternehmen Kosten der Integration von Informationssystemen strukturiert ermitteln können. Zudem erfolgt eine Erweiterung des Ilmenauer Integrationsmodells – einem Modell zur Bestimmung des Integrationsgrads von Informationssystemen – um eine Kostenkomponente

Recognition of a translocation motif in the regulator HpaA from Xanthomonas euvesicatoria is controlled by the type III secretion chaperone HpaB

The Gram-negative plant-pathogenic bacterium Xanthomonas euvesicatoria is the causal agent of bacterial spot disease in pepper and tomato plants. Pathogenicity of X. euvesicatoria depends on a type III secretion (T3S) system which translocates effector proteins into plant cells and is associated with an extracellular pilus and a translocon in the plant plasma membrane. Effector protein translocation is activated by the cytoplasmic T3S chaperone HpaB which presumably targets effectors to the T3S system. We previously reported that HpaB is controlled by the translocated regulator HpaA which binds to and inactivates HpaB during the assembly of the T3S system. In the present study, we show that translocation of HpaA depends on the T3S substrate specificity switch protein HpaC and likely occurs after pilus and translocon assembly. Translocation of HpaA requires the presence of a translocation motif (TrM) in the N-terminal region. The TrM consists of an arginine-and proline-rich amino acid sequence and is also essential for the in vivo function of HpaA. Mutation of the TrM allowed the translocation of HpaA in hpaB mutant strains but not in the wild-type strain, suggesting that the recognition of the TrM depends on HpaB. Strikingly, the contribution of HpaB to the TrM-dependent translocation of HpaA was independent of the presence of the C-terminal HpaB-binding site in HpaA. We propose that HpaB generates a recognition site for the TrM at the T3S system and thus restricts the access to the secretion channel to effector proteins. Possible docking sites for HpaA at the T3S system were identified by in vivo and in vitro interaction studies and include the ATPase HrcN and components of the predicted cytoplasmic sorting platform of the T3S system. Notably, the TrM interfered with the efficient interaction of HpaA with several T3S system components, suggesting that it prevents premature binding of HpaA. Taken together, our data highlight a yet unknown contribution of the TrM and HpaB to substrate recognition and suggest that the TrM increases the binding specificity between HpaA and T3S system components

Arbeitsbericht Nr. 2015-01, Januar 2015

Die deutsche Druckbranche ist geprägt von kleinen und mittelständischen Unternehmen (KMU). Wachsender Wettbewerb, immer individuellere Kundenwünsche bei sinkenden Auflagen und hohe Auftragsmanagementkosten tragen dazu bei, dass KMU sich zu Produktionsverbünden zusammenschließen. Um die gemeinsamen Wertschöpfungsprozesse in diesen Verbünden effizient gestalten zu können, sollten die Informationssysteme sowohl der kooperierenden Druckdienstleister als auch die ihrer Kunden adäquat integriert werden. Zur Ermittlung der unternehmensübergreifenden Integrationsbedarfe in Produktionsverbünden der Druckbranche wurde ein Modell entwickelt, welches die bei einer IT-Integration zu integrierenden Prozesse, Funktionen und Daten beschreibt. In der vorliegenden Arbeit werden durch Anwendung dieses Integrationsmodells verschiedene Integrationsoptionen für Druckdienstleister beschrieben und bewertet

Closure times of neurocranial sutures and synchondroses in Persian compared to Domestic Shorthair cats

Human-directed selective breeding has modified the phenotype of the modern Persian cat towards an extreme brachycephalic phenotype (‘peke-face’ Persian), which originates from a spontaneous mutation that first appeared in the 1950s in traditional Persian types. It was suggested that the peke-face phenotype results from pathologic skull development and might represent a craniosynostosis of the coronal sutures. We followed this hypothesis and investigated the time dependent status of the neurocranial sutures and synchondroses in an ontogenetic series of doll-faced and peke-faced Persian cats compared to Domestic Shorthair cats (DSHs). Cranial suture closure was assessed by examining an ontogenetic series of formalin-fixed head specimens (n = 55) and dry skulls (n = 32) using micro-computed tomography. Sagittal, metopic, coronal and lambdoid sutures as well as intersphenoidal, spheno-occipital and spheno-ethmoid synchondroses were examined. Logistic regression analysis was performed to test the global effect of age on suture closure within a group of peke-face Persians, doll-face Persians and DSHs and the 50% probability of having a closed suture was calculated and compared between groups. Age was a perfect predictor for the condition of the coronal sutures in peke-face Persians. Coronal sutures were found to be closed at 0–0.3 months. In doll-face and DSHs, coronal sutures were open throughout the lifetime with the exception of a few very old cats. Results of this study confirmed a coronal craniosynostosis that likely causes the extreme brachycephalic skull morphology in the peke-face Persian

Reliability and interobserver variability of a grading system of ventricular distension in dogs

IntroductionInternal hydrocephalus is the most common malformation of the central nervous system in dogs. Although the grades of ventricular distension have importance for long-term prognosis, there is no standard classification scheme describing the grade of the ventricular distension in dogs.Materials and methodsMagnetic resonance imaging (MRI) scans from 147 dogs of various breed, sex, skull conformation, and weight were reviewed retrospectively and blinded between three observers. Based on objectively assessable morphologic characteristics, the lateral cerebral ventricles were graded as normal, minimally, mildly, moderately, severely enlarged or end stage (grade 0 to grade 5), respectively. Evans’ index or the ventricle brain index was also measured in all animals. Interobserver agreement between a very experienced, experienced, and unexperienced person was evaluated by the Spearman coefficient and kappa tests. Additionally, correlation to the ventricle brain index was determined using the Spearman coefficient and F-tests.ResultsThe Spearman correlation coefficient reached a very strong correlation (r = 0.97) between the experienced and very experienced observer and a strong correlation (r = 0.91) between the very experienced and unexperienced observer. The kappa value revealed excellent interobserver agreement between the very experienced and experienced observers (weighted kappa 0.91) and moderate between the very experienced and unexperienced observers (weighted kappa 0.75). The ventricular-brain index correlated (r = 0.94, Spearman coefficient test) with the grading system, indicating that a more elevated ratio was related to a more advanced degree of ventricular enlargement. The interobserver agreement with regard to the grade between the neurologist in training and a board-certified neurologist was excellent and between the board-certified neurologist and general practitioner achieved lower values.ConclusionThe presented MRI-based grading of ventricular enlargement is a reliable and functional method for an objective grading of the ventricular system in dogs. Some experience in MRI and brain anatomy is needed for interpretation and grading

Volcanic jets, plumes, and collapsing fountains: evidence from large-scale experiments, with particular emphasis on the entrainment rate

The source conditions of volcanic plumes and collapsing fountains are investigated by means of large-scale experiments. In the experiments, gas-particle jets issuing from a cylindrical conduit are forced into the atmosphere at different mass flow rates. Dense jets (high particle volumetric concentration, e.g., C 0 > 0.01) generate collapsing fountains, whose height scales with the squared exit velocity. This is consistent with Bernoulli's equation, which is a good approximation if air entrainment is negligible. In this case, kinetic energy is transformed into potential energy without any significant loss by friction with the atmosphere. The dense collapsing fountain, on hitting the ground, generates an intense shear flow similar to a pyroclastic density current. Dilute hot jets (low particle volumetric concentration, e.g., C 0 3). © 2014 Springer-Verlag Berlin Heidelberg

HpaC Controls Substrate Specificity of the Xanthomonas Type III Secretion System

The Gram-negative bacterial plant pathogen Xanthomonas campestris pv. vesicatoria employs a type III secretion (T3S) system to inject bacterial effector proteins into the host cell cytoplasm. One essential pathogenicity factor is HrpB2, which is secreted by the T3S system. We show that secretion of HrpB2 is suppressed by HpaC, which was previously identified as a T3S control protein. Since HpaC promotes secretion of translocon and effector proteins but inhibits secretion of HrpB2, HpaC presumably acts as a T3S substrate specificity switch protein. Protein–protein interaction studies revealed that HpaC interacts with HrpB2 and the C-terminal domain of HrcU, a conserved inner membrane component of the T3S system. However, no interaction was observed between HpaC and the full-length HrcU protein. Analysis of HpaC deletion derivatives revealed that the binding site for the C-terminal domain of HrcU is essential for HpaC function. This suggests that HpaC binding to the HrcU C terminus is key for the control of T3S. The C terminus of HrcU also provides a binding site for HrpB2; however, no interaction was observed with other T3S substrates including pilus, translocon and effector proteins. This is in contrast to HrcU homologs from animal pathogenic bacteria suggesting evolution of distinct mechanisms in plant and animal pathogenic bacteria for T3S substrate recognition