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Nuclear displacement and fluorescence recovery after photobleaching (FRAP) assays to study division site placement and cytokinesis in fission yeast.
Methods in Cell Biology
Cytokinesis is an essential cellular event that completes the cell division cycle. It begins with the assembly of an actomyosin contractile ring that undergoes constriction concomitant with the septum formation to divide the cell in two. Placement of the septum at the right position is important to ensure fidelity of the division process. In fission yeast, the medially placed nucleus is a major spatial cue to position the site of division. In this chapter, we describe a simple synthetic biology-based approach to displace the nucleus and study the consequence on division site positioning. We also describe how to perform fluorescence recovery after photobleaching to follow the dynamics of cytokinetic proteins at defined time points by live-cell microscopy.
Actin Cytoskeleton/ultrastructure, Actomyosin/genetics, Actomyosin/ultrastructure, Cell Nucleus/genetics, Cell Nucleus/ultrastructure, Cytokinesis/genetics, Fluorescence Recovery After Photobleaching/methods, Microscopy/methods, Schizosaccharomyces/genetics, Schizosaccharomyces/ultrastructure, Cell division, Cytokinesis, Division site placement, Fission yeast Schizosaccharomyces pombe, Fluorescence recovery after photobleaching, FRAP, Live-cell microscopy, Nuclear displacement, Synthetic biology
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