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• W2101443143 abstract "Proteins with pleckstrin homology (PH) domains have been shown to be mostly involved in cytoskeletal signalling. We have identified a new protein in Dictyostelium discoideum, whose primary structure analysis reveals the presence of two PH domains along with additional functional domains like CH, RhoGEF and ArfGAP domains. Henceforth the protein is referred to as PhdB (Pleckstrin homology domain containing protein B). Monoclonal and polyclonal antibodies generated against PhdB recognise a 175 kDa protein throughout the developmental differentiation programme of D. discoideum. A GFP fusion protein containing the C-terminal PH domain of PhdB colocalises with F-actin at the pseudopods and is associated with the Triton X-100 insoluble F-actin cytoskeleton. The recombinant C-terminal domain of PhdB also binds to phosphoinositides and actin in vitro. To define the function of the protein we generated null cells lacking the PhdB protein by homologous recombination. The phenotype of the null cells reveals that the protein is necessary for normal cell aggregation during development as PhdB- cells showed a developmental delay and failed to form streams unlike wild type cells during chemotaxis upon a challenge with an external source of cAMP. During further analysis of the developmental defect we observed that the expression of the cell-cell adhesion molecule contact site A (csA) occurred independent of the formation of cell-cell contacts in PhdB- cells. The csA protein is a cAMP induced glycoprotein mediating EDTA-stable contacts. Furthermore, its expresion followed a time course similar to the one observed in wild type and normal levels of csA were reached. However, the presence of the csA did not allow the formation of strong aggregates by PhdB- cells. It therefore appears that the normal expression of csA which we observed during aggregation in PhdB- cells is independent of cell-cell adhesion. Taken together, the abnormal aggregation and the defect in streaming of PhdB- cells indicate that the protein is a component of cell-cell signalling during Dictyostelium aggregation." @default.
• W2101443143 created "2016-06-24" @default.
• W2101443143 creator A5058980704 @default.
• W2101443143 date "2004-01-01" @default.
• W2101443143 modified "2023-09-26" @default.
• W2101443143 title "Characterisation of PhdB, a pleckstrin homology domain containing protein in Dictyostelium discoideum" @default.
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