A New Methodology for the Quantification of In Vivo Cdc14 Phosphatase Activity.
The phosphatase Cdc14 has a pivotal function in the mitotic exit of Saccharomyces cerevisiae. During interphase, Cdc14 remains inactive in the nucleolus bound to the inhibitor Net1. Cdc14 activation occurs in the metaphase to anaphase transition and it is promoted by at least two signaling pathways called FEAR (CdcFourteen Early Anaphase Release) and MEN (Mitotic Exit Network). These two pathways act in parallel and target the phosphorylation of Net1, thus decreasing Net1 affinity for Cdc14. The activity of Cdc14 can be used as a readout to assay functional interactions of different components of the mitotic exit signaling pathways.
Cancer Epigenetics & Biology Program, IDIBELL,Hospitalet de Llobregat, Barcelona, Spain. equeralt@idibell.cat.
Ethel
E
Queralt
Molecular Biology Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, NY, 10065, USA. rodrigj5@mskcc.org.
José-Antonio
JA
Rodriguez-Rodriguez
D018929:Cell Culture Techniques / Q000379:methods; D018797:Cell Cycle Proteins / Q000032:analysis / Q000378:metabolism*; D057075:Enzyme Assays / Q000379:methods*; D047468:Immunoprecipitation / Q000379:methods*; D008938:Mitosis; D010766:Phosphorylation; D017027:Protein Tyrosine Phosphatases / Q000032:analysis / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis / Q000378:metabolism*; D015398:Signal Transduction
C074860:CDC14 protein, S cerevisiae; D018797:Cell Cycle Proteins; D029701:Saccharomyces cerevisiae Proteins; D017027:Protein Tyrosine Phosphatases
Assay; Cdc14; DiFMUP; Fluorescent; Immunoprecipitation; Mitotic exit; Net1; Phosphatase