Identification of an Aurora-A/PinsLINKER/ Dlg Spindle Orientation Pathway using Induced Cell Polarity in S2 Cells
Copyright policy )Identification of an Aurora-A/PinsLINKER/ Dlg Spindle Orientation Pathway using Induced Cell Polarity in S2 Cells
Asymmetric cell division is intensely studied because it can generate cellular diversity as well as maintain stem cell populations. Asymmetric cell division requires mitotic spindle alignment with intrinsic or extrinsic polarity cues, but mechanistic detail of this process is lacking. Here, we develop a method to construct cortical polarity in a normally unpolarized cell line and use this method to characterize Partner of Inscuteable (Pins; LGN/AGS3 in mammals) -dependent spindle orientation. We identify a previously unrecognized evolutionarily conserved Pins domain (Pins(LINKER)) that requires Aurora-A phosphorylation to recruit Discs large (Dlg; PSD-95/hDlg in mammals) and promote partial spindle orientation. The well-characterized Pins(TPR) domain has no function alone, but placing the Pins(TPR) in cis to the Pins(LINKER) gives dynein-dependent precise spindle orientation. This "induced cortical polarity" assay is suitable for rapid identification of the proteins, domains, and amino acids regulating spindle orientation or cell polarity.
- Howard Hughes Medical Institute United States
- Slovak Academy of Sciences Slovakia
- Neurosciences Institute United States
- National Academy of Sciences of Armenia Armenia
- University of Oregon United States
Biochemistry, Genetics and Molecular Biology(all), Tumor Suppressor Proteins, Cell Polarity, Dyneins, Cell Cycle Proteins, Spindle Apparatus, Protein Serine-Threonine Kinases, Prophase, Cell Line, Protein Structure, Tertiary, Drosophila melanogaster, Aurora Kinases, Animals, Drosophila Proteins, CELLBIO, Phosphorylation, Microtubule-Associated Proteins, Guanine Nucleotide Dissociation Inhibitors, Signal Transduction
Biochemistry, Genetics and Molecular Biology(all), Tumor Suppressor Proteins, Cell Polarity, Dyneins, Cell Cycle Proteins, Spindle Apparatus, Protein Serine-Threonine Kinases, Prophase, Cell Line, Protein Structure, Tertiary, Drosophila melanogaster, Aurora Kinases, Animals, Drosophila Proteins, CELLBIO, Phosphorylation, Microtubule-Associated Proteins, Guanine Nucleotide Dissociation Inhibitors, Signal Transduction
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