Measuring proteolysis in mitosis.
The targeted destruction of key regulators helps to drive the cell cycle. Here we describe a quantitative assay to measure destruction of different regulators in mitotic cells. This assay uses GFP-tagged substrates and time-lapse fluorescence microscopy of single cells to pinpoint the timing of destruction of different substrates at different stages in mitosis.
Lindon
Catherine
C
Department of Genetics, University of Cambridge, Cambridge, UK.
Di Fiore
Barbara
B
D000705:Anaphase / Q000502:physiology; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002877:Chromosomes, Human / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008677:Metaphase / Q000502:physiology; D008845:Microinjections; D008856:Microscopy, Fluorescence; D008938:Mitosis / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D008941:Spindle Apparatus / Q000502:physiology*; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism
D018797:Cell Cycle Proteins; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D044767:Ubiquitin-Protein Ligases