Analysis of the Localization of MEN Components by Live Cell Imaging Microscopy.
Mitotic exit is determined by multiple spatial and temporal cues from the spindle poles and the two compartments in a dividing yeast cell-the mother and the bud. These signals are ultimately integrated by the activation of the mitotic exit network (MEN) to promote persistent release of Cdc14 from the nucleolus. Live imaging analysis using fluorescent protein tags is invaluable to dissect this critical decision-making trigger. Here, we present protocols for routine yeast live cell microscopy applicable to this problem.
Department of Genetics, University of Cambridge, Downing Street, Cambridge, CB2 3EH, UK.
Zhiang
Z
Guo
Department of Genetics, University of Cambridge, Downing Street, Cambridge, CB2 3EH, UK. ms433@cam.ac.uk.
Marisa
M
Segal
D000705:Anaphase; D018797:Cell Cycle Proteins / Q000032:analysis*; D049452:Green Fluorescent Proteins / Q000032:analysis; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D008938:Mitosis; D061848:Optical Imaging / Q000379:methods; D017346:Protein Serine-Threonine Kinases / Q000032:analysis*; D017027:Protein Tyrosine Phosphatases / Q000032:analysis; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis*; D059010:Single-Cell Analysis / Q000379:methods*
C074860:CDC14 protein, S cerevisiae; D018797:Cell Cycle Proteins; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins; D017346:Protein Serine-Threonine Kinases; C081927:CDC5 protein, S cerevisiae; D017027:Protein Tyrosine Phosphatases
Cell population analysis; Digital image; Fluorescence microscopy; Fluorescent protein tags; Live cell imaging; Single cell analysis; Still imaging; Time lapse