Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P50583 (asymmetrical)
 12,197 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

This protocol describes an assay for the induction of asymmetrical cell division where the entire spindle is segregated into only one of the daughter cells. The procedure consists of four stages: (i) generation of asymmetrical monoasters by arresting cells in early mitosis with a kinesin Eg5 inhibitor; (ii) induction of cell division by microinjection of recombinant Mad1 protein or by the addition of a Cdk1 inhibitor; (iii) monitoring the division process by phase-contrast time-lapse microscopy; and (iv) processing for correlative immunofluorescence or correlative electron microscopy. This approach can be applied to determine the requirement for the mitotic spindle in organelle partitioning as well as to investigate the role of the monopolar spindle in cytokinesis. Moreover, the generated nucleus-lacking cytoplast provides an ideal environment to test the feasibility and activity of biological processes in the absence of genomic influence. The protocol takes 2-4 d to complete.
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PMID:Induction of asymmetrical cell division to analyze spindle-dependent organelle partitioning using correlative microscopy techniques. 1987 22

Phosphatase and tensin homologue (Pten) suppresses neoplastic growth by negatively regulating PI(3)K signalling through its phosphatase activity. To gain insight into the actions of non-catalytic Pten domains in normal physiological processes and tumorigenesis, we engineered mice lacking the PDZ-binding domain (PDZ-BD). Here, we show that the PDZ-BD regulates centrosome movement and that its heterozygous or homozygous deletion promotes aneuploidy and tumour formation. We found that Pten is recruited to pre-mitotic centrosomes in a Plk1-dependent fashion to create a docking site for protein complexes containing the PDZ-domain-containing protein Dlg1 (also known as Sap97) and Eg5 (also known as Kif11), a kinesin essential for centrosome movement and bipolar spindle formation. Docking of Dlg1-Eg5 complexes to Pten depended on Eg5 phosphorylation by the Nek9-Nek6 mitotic kinase cascade and Cdk1. PDZ-BD deletion or Dlg1 ablation impaired loading of Eg5 onto centrosomes and spindle pole motility, yielding asymmetrical spindles that are prone to chromosome missegregation. Collectively, these data demonstrate that Pten, through the Dlg1-binding ability of its PDZ-BD, accumulates phosphorylated Eg5 at duplicated centrosomes to establish symmetrical bipolar spindles that properly segregate chromosomes, and suggest that this function contributes to tumour suppression.
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PMID:Pten regulates spindle pole movement through Dlg1-mediated recruitment of Eg5 to centrosomes. 2724 Mar 20