Linking sister chromatid cohesion and apoptosis: role of Rad21.
Rad21 is one of the major cohesin subunits that holds sister chromatids together until anaphase, when proteolytic cleavage by separase, a caspase-like enzyme, allows chromosomal separation. We show that cleavage of human Rad21 (hRad21) also occurs during apoptosis induced by diverse stimuli. Induction of apoptosis in multiple human cell lines results in the early (4 h after insult) generation of 64- and 60-kDa carboxy-terminal hRad21 cleavage products. We biochemically mapped an apoptotic cleavage site at residue Asp-279 (D(279)) of hRad21. This apoptotic cleavage site is distinct from previously described mitotic cleavage sites. hRad21 is a nuclear protein; however, the cleaved 64-kDa carboxy-terminal product is translocated to the cytoplasm early in apoptosis before chromatin condensation and nuclear fragmentation. Overexpression of the 64-kDa cleavage product results in apoptosis in Molt4, MCF-7, and 293T cells, as determined by TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) and Annexin V staining, assaying of caspase-3 activity, and examination of nuclear morphology. Given the role of hRad21 in chromosome cohesion, the cleaved C-terminal product and its translocation to the cytoplasm may act as a nuclear signal for apoptosis. In summary, we show that cleavage of a cohesion protein and translocation of the C-terminal cleavage product to the cytoplasm are early events in the apoptotic pathway and cause amplification of the cell death signal in a positive-feedback manner.
Pati
Debananda
D
Texas Children's Cancer Center, Department of Pediatrics, Baylor College of Medicine, Houston, Texas 77030, USA. pati@bcm.tmc.edu
Zhang
Nenggang
N
Plon
Sharon E
SE
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D018797:Cell Cycle Proteins; D002478:Cells, Cultured; D002842:Chromatids / Q000378:metabolism*; D004260:DNA Repair; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors / Q000378:metabolism; D005047:Etoposide / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism*; D006801:Humans; D020287:In Situ Nick-End Labeling; D008938:Mitosis / Q000502:physiology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins / Q000378:metabolism*; D019384:Nucleic Acid Synthesis Inhibitors / Q000378:metabolism; D010455:Peptides / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D011453:Prostaglandins / Q000378:metabolism; D016415:Sequence Alignment; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D061945:Caspase Inhibitors; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D009687:Nuclear Proteins; D019384:Nucleic Acid Synthesis Inhibitors; D010455:Peptides; D010750:Phosphoproteins; D011453:Prostaglandins; C429022:RAD21 protein, human; D016159:Tumor Suppressor Protein p53; D005047:Etoposide; D020169:Caspases