Interaction between cell division proteins FtsE and FtsZ.

FtsE and FtsX, which are widely conserved homologs of ABC transporters and interact with each other, have important but unknown functions in bacterial cell division. Coimmunoprecipitation of Escherichia coli cell extracts revealed that a functional FLAG-tagged version of FtsE, the putative ATP-binding component, interacts with FtsZ, the bacterial tubulin homolog required to assemble the cytokinetic Z ring and recruit the components of the divisome. This interaction is independent of FtsX, the predicted membrane component of the ABC transporter, which has been shown previously to interact with FtsE. The interaction also occurred independently of FtsA or ZipA, two other E. coli cell division proteins that interact with FtsZ. In addition, FtsZ copurified with FLAG-FtsE. Surprisingly, the conserved C-terminal tail of FtsZ, which interacts with other cell division proteins, such as FtsA and ZipA, was dispensable for interaction with FtsE. In support of a direct interaction with FtsZ, targeting of a green fluorescent protein (GFP)-FtsE fusion to Z rings required FtsZ, but not FtsA. Although GFP-FtsE failed to target Z rings in the absence of ZipA, its localization was restored in the presence of the ftsA* bypass suppressor, indicating that the requirement for ZipA is indirect. Coexpression of FLAG-FtsE and FtsX under certain conditions resulted in efficient formation of minicells, also consistent with an FtsE-FtsZ interaction and with the idea that FtsE and FtsX regulate the activity of the divisome.

Genetic Control of Plant Development by Overriding a Geometric Division Rule

Formative cell divisions are critical for multicellular patterning. In the early plant embryo, such divisions follow from orienting the division plane. A major unanswered question is how division plane orientation is genetically controlled, and in particular whether this relates to cell geometry. We have generated a complete 4D map of early Arabidopsis embryogenesis and used computational analysis to demonstrate that several divisions follow a rule that uses the smallest wall area going through the center of the cell. In other cases, however, cell division clearly deviates from this rule, which invariably leads to asymmetric cell division. By analyzing mutant embryos and through targeted genetic perturbation, we show that response to the hormone auxin triggers a deviation from the ``shortest wall'' rule. Our work demonstrates that a simple default rule couples division orientation to cell geometry in the embryo and that genetic regulation can create patterns by overriding the default rule.

The Caenorhabditis elegans histone hairpin-binding protein is required for core histone gene expression and is essential for embryonic and postembryonic cell division.

As in all metazoans, the replication-dependent histone genes of Caenorhabditis elegans lack introns and contain a short hairpin structure in the 3' untranslated region. This hairpin structure is a key element for post-transcriptional regulation of histone gene expression and determines mRNA 3' end formation, nuclear export, translation and mRNA decay. All these steps contribute to the S-phase-specific expression of the replication-dependent histone genes. The hairpin structure is the binding site for histone hairpin-binding protein that is required for hairpin-dependent regulation. Here, we demonstrate that the C. elegans histone hairpin-binding protein gene is transcribed in dividing cells during embryogenesis and postembryonic development. Depletion of histone hairpin-binding protein (HBP) function in early embryos using RNA-mediated interference leads to an embryonic-lethal phenotype brought about by defects in chromosome condensation. A similar phenotype was obtained by depleting histones H3 and H4 in early embryos, indicating that the defects in hairpin-binding protein-depleted embryos are caused by reduced histone biosynthesis. We have confirmed this by showing that HBP depletion reduces histone gene expression. Depletion of HBP during postembryonic development also results in defects in cell division during late larval development. In addition, we have observed defects in the specification of vulval cell fate in animals depleted for histone H3 and H4, which indicates that histone proteins are required for cell fate regulation during vulval development.

Sendschreiben an die Redakteure der in Leipzig erscheinenden Deutschen Kriegerzeitung : Wilucki, Oberlieutenant im 2. Linien-Inf.-Regt., Müller, Oberlieutenant in der Inf.-Garde-Division, Erdmannsdorf, Lieutenant im Leib-Inf.-Regt., v. Zychlinski, Lieutenant im Leib.-Inf.-Regt., Andrée, Lieutenant im Ingenieur-Corps

Boberach: Der Aufsatz "Wie denken Preußens Offiziere von einem deutschen Heere?" hat das Offizierskorps verletzt, daß sich gegen jede Mediatisierung Preußens wendet, weil Preußens Heer allein Deutschland vor dem Untergang retten kann

Kundmachung: In Folge einer Mittheilung des General-Commando's kann die Artillerie-Division ...

Welsch (Projektbearbeiter): Bekanntmachung eines Eisenbahnmilitärtransports von Prag über Wien nach Oberitalien

Nationalgarde 1. Cavallerie-Division: Vom Commando der 1. Nationalgarde-Cavallerie-Division wird hiemit der Befehl ertheilt ...

Welsch (Projektbearbeiter): Befehl an die Züge [Unterabteilungen der Truppenkörper] aus der Innen- sowie der Leopoldstadt, sich auf Dauer in der Stallburg zu versammeln

Genetic and functional analyses of cell division proteins FtsZ and FtsA in Escherichia coli

In the current model for bacterial cell division, the FtsZ protein forms a ring that marks the division plane, creating a cytoskeletal framework for the subsequent action of other essential division proteins such as FtsA and ZipA. The putative protein complex ultimately generates the division septum. The essential cell division protein FtsZ is a functional and structural homolog of eukaryotic tubulin, and like tubulin, FtsZ hydrolyzes GTP and self-assembles into protein filaments in a strictly GTP-dependent manner. FtsA shares sequence similarity with members of the ATPase superfamily that include actin, but its actual function remains unknown. To test the division model and elucidate functions of the division proteins, this dissertation primarily focuses on the analysis of FtsZ and FtsA in Escherichia coli. ^ By tagging with green fluorescent protein, we first demonstrated that FtsA also exhibits a ring-like structure at the potential division site. The localization of FtsA was dependent on functional FtsZ, suggesting that FtsA is recruited to the septum by the FtsZ ring. In support of this idea, we showed that FtsA and FtsZ directly interact. Using a novel E. coli in situ assay, we found that the FtsA-FtsZ interaction appears to be species-specific, although an interspecies interaction could occur between FtsA and FtsZ proteins from two closely related organisms. In addition, mutagenesis of FtsA revealed that no single domain is solely responsible for its septal localization or interaction with FtsZ. To explore the function of FtsA, we purified FtsA protein and demonstrated that it has ATPase activity. Furthermore, purified FtsA stimulates disassembly of FtsZ polymers in a sedimentation assay but does not affect GTP hydrolysis of FtsZ. This result suggests that in the cell, FtsA may function similarly in regulating dynamic instability of the FtsZ ring during the cell division process. ^ To elucidate the structure-function relationship of FtsZ, we carried out thorough genetic and functional analyses of the mutagenized FtsZ derivatives. Our results indicate that the conserved N-terminal domain of FtsZ is necessary and sufficient for FtsZ self-assembly and localization. Moreover, we discovered a critical role for an extreme C-terminal domain of FtsZ that consists of only 12 residues. Truncated FtsZ derivatives lacking this domain, though able to polymerize and localize, are defective in ring formation in vivo as well as interaction with FtsA and ZipA. Alanine scanning mutagenesis of this region pinpointed at least five residues necessary for the function of FtsZ. Studies of protein levels and protein-protein interactions suggested that these residues may be involved in regulating protein stability and/or FtsZ-FtsA interactions. Interestingly, two of the point mutants exhibited dominant-negative phenotypes. ^ In summary, results from this thesis work have provided additional support for the division machinery model and will contribute to a better understanding of the coordinate functions of FtsA and FtsZ in the cell division process. ^

Temporal and spatial regulation of cell division by the nucleoid in Escherichia coli

Selection of division sites and coordination of cytokinesis with other cell cycle events are critical for every organism to proliferate. In E. coli, the nucleoid is proposed to exclude division from the site of the chromosome (nucleoid occlusion model). We studied the effect of the nucleoid on timing and placement of cell division. An early cell division protein, FtsZ, was used to follow development of the division septum. FtsZ forms a ring structure (Z ring) at potential division sites. The dynamics of Z ring was visualized in live cells by fusing FtsZ with a green fluorescent protein (GFP). Emanating FtsZ-GFP polymers from the constricted septum or aggregates in daughter cells were also observed, probably representing the FtsZ depolymerization and immature FtsZ nucleation processes. We next examined the nucleoid occlusion model. Mutants carrying abnormally positioned chromosomes were employed. In chromosomal partition mutants, replicated chromosomes cannot segregate. The Z ring was excluded from midcell to the edge of the nucleoid. This negative effect of nucleoids was further confirmed in replication deficient dnaA mutants, in which only a single chromosome is present in the cell center. These results suggest that the nucleoid, replicating or not, inhibits division in the area where the chromosome occupies. In addition, increasing the level of FtsZ does not overcome nucleoid inhibition. Interestingly in anucleate cells produced by both mutants, the Z ring was localized in the central part of the cell, which indicates that the nucleoid is not required for FtsZ assembly. Relaxation of chromosomes by reducing the gyrase activity or disruption of protein translation/translocation did not abolish the division inhibition capacity of the nucleoid. However, preventing transcription did compromise the nucleoid occlusion effect, leading to formation of multiple FtsZ rings above the nucleoid. In summary, we demonstrate that nucleoids negatively regulate the timing and position of division by inhibiting FtsZ assembly at unselected sites. Relief of this inhibition at midcell is coincident with the completion of DNA replication. On the other hand, FtsZ assembly does not require the nucleoid. ^

Festrede, bei der Fahnen- und Estandarten-Weihe der vier Stadtbezirke und des XII. Bezirkes, so wie der National-Garde-Cavallerie-Division - Fest-Rede, bei der Fahnen- und Estandarten-Weihe der vier Stadtbezirke und des XII. Bezirkes, so wie der National-Garde-Cavallerie-Division : am 24. September 1848

gesprochen von Joseph Holzapfel

Identifying the roles of the Escherichia coli FtsZ-associated proteins by minimizing the requirements for division

Formation of the FtsZ ring (Z ring) in Escherichia coli is the first step in assembly of the divisome, a molecular machine composed of 14 known proteins which are all required for cell division. Although the biochemical functions of most divisome proteins are unknown, several of these have overlapping roles in ensuring that the Z ring assembles at the cytoplasmic membrane and is active. ^ We identified a single amino acid change in FtsA, R286W, renamed FtsA*, that completely bypasses the requirement for ZipA in cell division. This and other data suggest that FtsA* is a hyperactive form of FtsA that can replace the multiple functions normally assumed by ZipA, which include stabilization of Z rings, recruitment of downstream cell division proteins, and anchoring the Z ring to the membrane. This is the first example of complete functional replacement of an essential prokaryotic cell division protein by another. ^ Cells expressing ftsA* with a complete deletion of ftsK are viable and divide, although many of these ftsK null cells formed multiseptate chains, suggesting a role in cell separation for FtsK. In addition, strains expressing extra ftsAZ, ftsQ, ftsB, zipA or ftsN, were also able to survive and divide in the absence of ftsK. The cytoplasmic and transmembrane domains of FtsQ were sufficient to allow viability and septum formation to ftsK deleted strains. These findings suggest that FtsK is normally involved in stabilizing the divisome and shares functional overlap with other cell division proteins. ^ As well as permitting the removal of other divisome components, the presence of FtsA* in otherwise wild-type cells accelerated Z-ring assembly, which resulted in a significant decrease in the average length of cells. In support of its role in Z-ring stability, FtsA* suppressed the cell division inhibition caused by overexpressing FtsZ. FtsA* did not affect FtsZ turnover within the Z ring as measured by fluorescence recovery after photobleaching. Turnover of FtsA* in the ring was somewhat faster than wild-type FtsA. Yeast two-hybrid data suggest that FtsA* has an increased affinity for FtsZ relative to wild-type FtsA. These results indicate that FtsA* interacts with FtsZ more strongly, and its enhancement of Z ring assembly may explain why FtsA* can permit survival of cells lacking ZipA or FtsK.^

The results of a barrier analysis on complementary feeding practices among mothers in the Central River Division of the Gambia

Currently, the barriers to appropriate infant feeding practices are largely unknown in the Central River Division of the Gambia. A questionnaire was developed and implemented by a local Non Governmental Organization (NGO), the Gambia Food and Nutrition Agency, in order to gain more information and ultimately to improve the child mortality rate of the country. There were two participant groups: 88 Doers who are women who had adopted the appropriate complementary feeding practice guidelines as defined by the World Health Organization and 87 Non Doers who are women who had in some way strayed from the appropriate complementary feeding practice guidelines. The questionnaire included aspects of the Health Belief Model which can be used in the development of a future intervention. The Yes/No questions were analyzed using the Chi-square statistical method and the open-ended questions used a descriptive analysis method of evaluation. The constructs for perceived susceptibility, perceived action efficacy, perceived self efficacy, cues for action and perception of divine showed significant differences between the Doers and the Non Doers (p<0.05). The descriptive analysis revealed that both participant groups had a limited understanding of the preventative qualities of the adoption of appropriate complementary feeding practices. The women in both of groups also showed a strong perception of divine will. Women in the Central River Division perceive their husband and in-laws to be the most influential in the decision-making process regarding infant feeding practices. Recommendations for future interventions must acknowledge the importance and influence of the community surrounding the women in their adoption of the appropriate infant feeding practices. It would also be important to educate women about of the specific guidelines of the appropriate complementary feeding practices, specifically the delay in early initiation of complementary feeding. The results of this barrier analysis provide useful information to plan and implement an effective intervention to improve the child mortality rate in the Gambia. ^

THE EFFECTS OF COST-SHARING ON USE OF SERVICES AND CONSUMER CHARACTERISTICS IN A STATE'S CRIPPLED CHILDREN'S DIVISION

The present study analyzed some of the effects of imposing a cost-sharing requirement on users of a state's health service program. The study population consisted of people who were in diagnosed medical need and included, but was not limited to, people in financial need.^ The purpose of the study was to determine if the cost-sharing requirement had any detrimental effects on the service population. Changes in the characteristics of service consumers and in utilization patterns were analyzed using time-series techniques and pre-post policy comparisons.^ The study hypotheses stated that the distribution of service provided, diagnoses serviced, and consumer income levels would change following the cost-sharing policy.^ Analysis of data revealed that neither the characteristics of service users (income, race, sex, etc.) nor services provided by the program changed significantly following the policy. The results were explainable in part by the fact that all of the program participants were in diagnosed medical need. Therefore, their use of "discretionary" or "less necessary" services was limited.^ The study's findings supported the work of Joseph Newhouse, Charles Phelps, and others who have contended that necessary service use would not be detrimentally affected by reasonable cost-sharing provisions. These contentions raise the prospect of incorporating cost-sharing into programs such as Medicaid, which, at this writing, do not demand any consumer payment for services.^ The study concluded with a discussion of the cost-containment problem in health services. The efficacy of cost-sharing was considered relative to other financing and reimbursement strategies such as HMO's, self-funding, and reimbursement for less costly services and places of service. ^

The spatial and linguistic division of the city of Malaga

The city of Malaga underwent considerable growth in the 19th and 20th centuries. The territorial expansion paired with a massive influx of immigrants occurred in three waves and as a consequence the city of Malaga remains divided into three different parts up to today. The differences between these three neighbourhoods of the city lie in the type of houses, different cultural and industrial activities, socioeconomic level, and very interestingly, also in speech. Thus, the aim of this study is an examination of the interrelation between speech (phonetic features) and urban space in Malaga. A combination of quantitative and qualitative analysis was used, based on two types of data: 1) production data stemming from recordings of 120 speakers; 2) perception data (salience, estimated frequency of use, attitude, spatial and social perception, imitation) which was collected from several surveys with 120 participants each. Results show that the speech production data divides the city of Malaga clearly into three different parts. This tripartition is confirmed by the analysis of the perception data. Moreover, the habitants of these three areas are perceived as different social types, to whom a range of social features is attributed. That is, certain linguistic features, the different neighbourhoods of the city and the social characteristics associated with them are undergoing a process of indexicalization and iconization. As a result, the linguistic features in question function as identity markers on the intraurban level.

Modeling of the Division Point of Different Propagation Mechanisms in the Near-Region Within Arched Tunnels

An accurate characterization of the near-region propagation of radio waves inside tunnels is of practical importance for the design and planning of advanced communication systems. However, there has been no consensus yet on the propagation mechanism in this region. Some authors claim that the propagation mechanism follows the free space model, others intend to interpret it by the multi-mode waveguide model. This paper clarifies the situation in the near-region of arched tunnels by analytical modeling of the division point between the two propagation mechanisms. The procedure is based on the combination of the propagation theory and the three-dimensional solid geometry. Three groups of measurements are employed to verify the model in different tunnels at different frequencies. Furthermore, simplified models for the division point in five specific application situations are derived to facilitate the use of the model. The results in this paper could help to deepen the insight into the propagation mechanism within tunnel environments.

Ultra-High Efficiency, High-Concentration PV System Based On Spectral Division Between GaInP/GaInAs/Ge And BPC Silicon Cells

A novel HCPV nonimaging concentrator concept with high concentration (>500×) is presented. It uses the combination of a commercial concentration GaInP∕GaInAs∕Ge 3J cell and a concentration Back‐Point‐Contact (BPC) concentration silicon cell for efficient spectral utilization, and external confinement techniques for recovering the 3J cell′s reflection. The primary optical element (POE) is a flat Fresnel lens and the secondary optical element (SOE) is a free‐form RXI‐type concentrator with a band‐pass filter embedded it, both POE and SOE performing Köhler integration to produce light homogenization. The band‐pass filter sends the IR photons in the 900–1200 nm band to the silicon cell. Computer simulations predict that four‐terminal terminal designs could achieve ∼46% added cell efficiencies using commercial 39% 3J and 26% Si cells. A first proof‐of concept receiver prototype has been manufactured using a simpler optical architecture (with a lower concentration, ∼ 100× and lower simulated added efficiency), and experimental measurements have shown up to 39.8% 4J receiver efficiency using a 3J with peak efficiency of 36.9%