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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Investigating prokaryotic communities : group activities and physiological heterogeneity

Wessel, Aimee Katherine 02 March 2015 (has links)
Bacterial communities engage in social activities, exhibiting behaviors such as communicating with small signaling molecules (quorum sensing [QS]) and building antibiotic-resistant biofilms. The opportunistic human pathogen Pseudomonas aeruginosa produces both freely diffusible QS molecules, as well as a QS molecule that is packaged or transported across cell membranes via the production of outer membrane vesicles. Despite the ubiquity of vesicle production in bacteria, the mechanism of outer membrane vesicle production has not been fully elucidated. In addition, most of our understanding of QS and biofilm formation arises from in vitro studies of bacterial communities containing large numbers of cells, often with greater than 10⁸ bacteria. However, many bacterial communities are comprised of small, densely packed aggregates of cells (≤10⁵ bacteria), and it is unclear how group behaviors and chemical interactions take place in densely packed, small populations. This dissertation has two main goals: i) to provide insights into the mechanism of bacterial membrane vesicle production, and ii) to understand how population size and the spatial distribution of cells affect cell-cell interactions and the nutritional microenvironment within a small (≤10⁵ bacteria) prokaryotic community. / text
2

Structural and Functional Characterization of the Soluble Cell Adhesion Molecule DdCAD-1in Dictyostelium discoideum

Sriskanthadevan, Shrivani 31 August 2011 (has links)
The cadA gene in Dictyostelium encodes a unique Ca2+-dependent cell adhesion molecule DdCAD-1. It is synthesized as a soluble protein in the cytoplasm and then transported to the plasma membrane by contractile vacuoles. The solution structures of Ca2+-free and Ca2+-bound DdCAD-1 reveals that it contains two β-sandwich domains, belonging to the βγ-crystallin and immunoglobulin fold classes, respectively. Whereas the N-terminal domain has a major role in homophilic binding, the C-terminal domain tethers the protein to the cell membrane. Although hydrophobic interactions constitute the major force for adhesion, electrostatic interactions may act as a ‘switch’ to regulate the homophilic binding by a change in electrostatic potential caused by the binding of Ca2+ to the three binding sites. To further investigate DdCAD-1 transport, DdCAD-1-GFP fusion proteins were expressed in cadA-null cells. Time-lapse microscopy revealed that DdCAD-1 was imported by invagination of the contractile vacuole membrane. The N-terminal, C-terminal domains, and two of the three Ca2+-binding site mutant forms of DdCAD-1 failed to enter the contractile vacuole, suggesting that Ca2+-binding and the integrity of DdCAD-1 are required for import. Indeed, proteins with altered conformation failed to enter the contractile vacuole, indicating that the import signal is integrated in the three-dimensional structure of DdCAD-1. Finally, we describe how the cadA gene acts as a single-gene green beard. In chimera experiments, cells expressing DdCAD-1 were more likely to form fruiting bodies than cadA-null cells on soil plates. Here cadA behaved as a single gene green beard. However, cadA exhibited anti-green beard behaviour on non-nutrient agar plates. Wild-type cells differentiated mostly into prestalk cells and eventually died, whereas the cadA-null cells survived as spores. DdCAD-1 was enriched in cell-cell contact regions of anterior cells, while it was mostly localized in the cytoplasm of posterior cells. The presence of DdCAD-1 on the cell surface of prestalk cells is crucial for cell sorting, which in turn explain the anti-green beard effect observed in chimeras containing cadA+ and cadA- cells. These observations demonstrate that DdCAD-1 plays a direct role in cell sorting through differential cell-cell adhesion which results from the differential distribution of DdCAD-1.
3

Structural and Functional Characterization of the Soluble Cell Adhesion Molecule DdCAD-1in Dictyostelium discoideum

Sriskanthadevan, Shrivani 31 August 2011 (has links)
The cadA gene in Dictyostelium encodes a unique Ca2+-dependent cell adhesion molecule DdCAD-1. It is synthesized as a soluble protein in the cytoplasm and then transported to the plasma membrane by contractile vacuoles. The solution structures of Ca2+-free and Ca2+-bound DdCAD-1 reveals that it contains two β-sandwich domains, belonging to the βγ-crystallin and immunoglobulin fold classes, respectively. Whereas the N-terminal domain has a major role in homophilic binding, the C-terminal domain tethers the protein to the cell membrane. Although hydrophobic interactions constitute the major force for adhesion, electrostatic interactions may act as a ‘switch’ to regulate the homophilic binding by a change in electrostatic potential caused by the binding of Ca2+ to the three binding sites. To further investigate DdCAD-1 transport, DdCAD-1-GFP fusion proteins were expressed in cadA-null cells. Time-lapse microscopy revealed that DdCAD-1 was imported by invagination of the contractile vacuole membrane. The N-terminal, C-terminal domains, and two of the three Ca2+-binding site mutant forms of DdCAD-1 failed to enter the contractile vacuole, suggesting that Ca2+-binding and the integrity of DdCAD-1 are required for import. Indeed, proteins with altered conformation failed to enter the contractile vacuole, indicating that the import signal is integrated in the three-dimensional structure of DdCAD-1. Finally, we describe how the cadA gene acts as a single-gene green beard. In chimera experiments, cells expressing DdCAD-1 were more likely to form fruiting bodies than cadA-null cells on soil plates. Here cadA behaved as a single gene green beard. However, cadA exhibited anti-green beard behaviour on non-nutrient agar plates. Wild-type cells differentiated mostly into prestalk cells and eventually died, whereas the cadA-null cells survived as spores. DdCAD-1 was enriched in cell-cell contact regions of anterior cells, while it was mostly localized in the cytoplasm of posterior cells. The presence of DdCAD-1 on the cell surface of prestalk cells is crucial for cell sorting, which in turn explain the anti-green beard effect observed in chimeras containing cadA+ and cadA- cells. These observations demonstrate that DdCAD-1 plays a direct role in cell sorting through differential cell-cell adhesion which results from the differential distribution of DdCAD-1.

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