The 14-3-3 protein rad24p modulates function of the cdc14p family phosphatase clp1p/flp1p in fission yeast.
Schizosaccharomyces pombe cells divide through the use of an actomyosin-based contractile ring. In response to perturbation of the actomyosin ring, S. pombe cells delay in a "cytokinesis-competent" state characterized by continuous repair and maintenance of the actomyosin ring and a G2 delay. This checkpoint mechanism requires the function of the Cdc14p-family phosphatase Clp1p/Flp1p and the septation initiation network (SIN). In response to cytokinetic defects, Clp1p, normally nucleolar in interphase, is retained in the cytoplasm until completion of cell division in a SIN-dependent manner. Here, we show that a phosphorylated form of Clp1p binds the 14-3-3 protein Rad24p and is retained in the cytoplasm in a Rad24p-dependent manner in response to cytokinesis defects. This physical interaction depends on the function of the SIN component, Sid2p. In the absence of Rad24p, cells are unable to maintain SIN signaling and lose viability upon mild cytokinetic stress. The requirement of Rad24p in this checkpoint is bypassed by ectopic activation of the SIN. Furthermore, SIN-dependent nuclear exclusion of Clp1p is dependent on Rad24p function. We conclude that Rad24p-mediated cytoplasmic retention of Clp1p/Flp1p is important for cell viability upon stress to the division apparatus.
Mishra
Mithilesh
M
Cell Division Laboratory, Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604.
Karagiannis
Jim
J
Sevugan
Mayalagu
M
Singh
Pritpal
P
Balasubramanian
Mohan K
MK
D000205:Actomyosin / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism*; D048749:Cytokinesis / Q000502:physiology*; D003593:Cytoplasm / Q000378:metabolism; D005982:Glutathione Transferase; D049452:Green Fluorescent Proteins; D015151:Immunoblotting; D047468:Immunoprecipitation; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D012568:Schizosaccharomyces; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*
D018797:Cell Cycle Proteins; D047908:Intracellular Signaling Peptides and Proteins; D029702:Schizosaccharomyces pombe Proteins; C501114:rad24 protein, S pombe; D049452:Green Fluorescent Proteins; D000205:Actomyosin; D005982:Glutathione Transferase; C439269:Clp1 protein, S. pombe; D017027:Protein Tyrosine Phosphatases