Caspase cleavage of MST1 promotes nuclear translocation and chromatin condensation

MST1, mammalian STE20-like kinase 1, is a serine/threonine kinase that is cleaved and activated by caspases during apoptosis. MST1 is capable of inducing apoptotic morphological changes such as chromatin condensation upon overexpression. In this study, we show that MST1 contains two functional nuclear export signals (NESs) in the C-terminal domain, which is released from the N-terminal kinase domain upon caspase-mediated cleavage. Full-length MST1 is excluded from the nucleus and localized to the cytoplasm. However, either truncation of the C-terminal domain, point mutation of the two putative NESs, or treatment with leptomycin B, an inhibitor of the NES receptor, results in nuclear localization of MST1. Staurosporine treatment induces chromatin condensation, MST1 cleavage, and nuclear translocation. Staurosporine-induced chromatin condensation is partially inhibited by expressing a kinase-negative mutant of MST1, suggesting an important role of MST1 in this process. Significantly, MST1 is more efficient at inducing chromatin condensation when it is constitutively localized to the nucleus by mutation of its NESs. Moreover, inhibition of MST1 nuclear translocation by mutation of its cleavage sites reduces its ability to induce chromatin condensation. Taken together, these results suggest that truncation of the C-terminal domain of MST1 by caspases may result in translocation of MST1 into the nucleus, where it promotes chromatin condensation.

[1]  E. Krebs,et al.  Both Phosphorylation and Caspase-mediated Cleavage Contribute to Regulation of the Ste20-like Protein Kinase Mst1 during CD95/Fas-induced Apoptosis* , 2001, The Journal of Biological Chemistry.

[2]  E. Sahai,et al.  Membrane blebbing during apoptosis results from caspase-mediated activation of ROCK I , 2001, Nature Cell Biology.

[3]  J. Bertoglio,et al.  Caspase-3-mediated cleavage of ROCK I induces MLC phosphorylation and apoptotic membrane blebbing , 2001, Nature Cell Biology.

[4]  E. Nishida,et al.  Polo-like kinase 1 phosphorylates cyclin B1 and targets it to the nucleus during prophase , 2001, Nature.

[5]  S. Kornbluth,et al.  Combinatorial Control of Cyclin B1 Nuclear Trafficking through Phosphorylation at Multiple Sites* , 2001, The Journal of Biological Chemistry.

[6]  Anthony J. Muslin,et al.  14-3-3 proteins: regulation of subcellular localization by molecular interference. , 2000, Cellular signalling.

[7]  Tak W. Mak,et al.  Two Distinct Pathways Leading to Nuclear Apoptosis , 2000, The Journal of experimental medicine.

[8]  V. Ferrans,et al.  Modulation of retinoid signalling through NGF-induced nuclear export of NGFI-B , 2000, Nature Cell Biology.

[9]  B. Henderson,et al.  A comparison of the activity, sequence specificity, and CRM1-dependence of different nuclear export signals. , 2000, Experimental cell research.

[10]  Guido Kroemer,et al.  Mitochondrio‐nuclear translocation of AIF in apoptosis and necrosis , 2000, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[11]  M. Watabe,et al.  Activation of MST/Krs and c-Jun N-terminal Kinases by Different Signaling Pathways during Cytotrienin A-induced Apoptosis* , 2000, The Journal of Biological Chemistry.

[12]  M. Rudnicki,et al.  Caspase 3 Cleavage of the Ste20-Related Kinase SLK Releases and Activates an Apoptosis-Inducing Kinase Domain and an Actin-Disassembling Region , 2000, Molecular and Cellular Biology.

[13]  G. Rodan,et al.  Bisphosphonates Act Directly on the Osteoclast to Induce Caspase Cleavage of Mst1 Kinase during Apoptosis , 1999, The Journal of Biological Chemistry.

[14]  T. Tan,et al.  Caspase-mediated cleavage and functional changes of hematopoietic progenitor kinase 1 (HPK1) , 1999, Oncogene.

[15]  D. Nicholson,et al.  Caspase structure, proteolytic substrates, and function during apoptotic cell death , 1999, Cell Death and Differentiation.

[16]  M. Watabe,et al.  Requirement of protein kinase (Krs/MST) activation for MT-21-induced apoptosis , 1999, Oncogene.

[17]  N. Imamoto,et al.  Acinus is a caspase-3-activated protein required for apoptotic chromatin condensation , 1999, Nature.

[18]  S. Nagata,et al.  Apoptotic nuclear morphological change without DNA fragmentation , 1999, Current Biology.

[19]  L. Ellerby,et al.  Release of caspase-9 from mitochondria during neuronal apoptosis and cerebral ischemia. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[20]  S. Korsmeyer,et al.  Cell Death in Development , 1999, Cell.

[21]  P. Russell,et al.  Nuclear localization of Cdc25 is regulated by DNA damage and a 14-3-3 protein , 1999, Nature.

[22]  J. Boyer,et al.  Caspase-3 controls both cytoplasmic and nuclear events associated with Fas-mediated apoptosis in vivo. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[23]  H. Kakeya,et al.  Caspase-mediated activation of a 36-kDa myelin basic protein kinase during anticancer drug-induced apoptosis. , 1998, Cancer research.

[24]  K. Arai,et al.  Signalling into the T-cell nucleus: NFAT regulation. , 1998, Cellular signalling.

[25]  Y. Lazebnik,et al.  Caspases: enemies within. , 1998, Science.

[26]  M. Yanagida,et al.  Leptomycin B inhibition of signal-mediated nuclear export by direct binding to CRM1. , 1998, Experimental cell research.

[27]  P. Li,et al.  The 40-kDa subunit of DNA fragmentation factor induces DNA fragmentation and chromatin condensation during apoptosis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[28]  J. Moore,et al.  Control of cyclin B1 localization through regulated binding of the nuclear export factor CRM1. , 1998, Genes & development.

[29]  E. Nishida,et al.  Proteolytic activation of MST/Krs, STE20-related protein kinase, by caspase during apoptosis , 1998, Oncogene.

[30]  V. Cryns,et al.  Proteases to die for. , 1998, Genes & development.

[31]  D. Templeton,et al.  Fas-induced proteolytic activation and intracellular redistribution of the stress-signaling kinase MEKK1. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[32]  E. Krebs,et al.  Caspase‐mediated activation and induction of apoptosis by the mammalian Ste20‐like kinase Mst1 , 1998, The EMBO journal.

[33]  C. Widmann,et al.  MEK Kinase 1, a Substrate for DEVD-Directed Caspases, Is Involved in Genotoxin-Induced Apoptosis , 1998, Molecular and Cellular Biology.

[34]  S. Lowe,et al.  Essential contribution of caspase 3/CPP32 to apoptosis and its associated nuclear changes. , 1998, Genes & development.

[35]  S. Nagata,et al.  Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis , 1998, Nature.

[36]  S. Nagata,et al.  A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD , 1998, Nature.

[37]  G. Salvesen,et al.  The Regulation of Anoikis: MEKK-1 Activation Requires Cleavage by Caspases , 1997, Cell.

[38]  G M Bokoch,et al.  Membrane and morphological changes in apoptotic cells regulated by caspase-mediated activation of PAK2. , 1997, Science.

[39]  Erich A. Nigg,et al.  Nucleocytoplasmic transport: signals, mechanisms and regulation , 1997, Nature.

[40]  Xiaodong Wang,et al.  DFF, a Heterodimeric Protein That Functions Downstream of Caspase-3 to Trigger DNA Fragmentation during Apoptosis , 1997, Cell.

[41]  E. Nishida,et al.  Interaction of MAP kinase with MAP kinase kinase: its possible role in the control of nucleocytoplasmic transport of MAP kinase , 1997, The EMBO journal.

[42]  H. de Thé,et al.  Arsenic-induced PML targeting onto nuclear bodies: implications for the treatment of acute promyelocytic leukemia. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[43]  R. Erikson,et al.  Newly identified stress-responsive protein kinases, Krs-1 and Krs-2. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[44]  J. Chernoff,et al.  The Ste20-like Protein Kinase, Mst1, Dimerizes and Contains an Inhibitory Domain* , 1996, The Journal of Biological Chemistry.

[45]  J. Richie,et al.  UV irradiation-induced apoptosis leads to activation of a 36-kDa myelin basic protein kinase in HL-60 cells. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[46]  J. Chernoff,et al.  Cloning and characterization of a member of the MST subfamily of Ste20-like kinases. , 1995, Gene.

[47]  D. Fujiwara,et al.  Leptomycin B targets a regulatory cascade of crm1, a fission yeast nuclear protein, involved in control of higher order chromosome structure and gene expression. , 1994, The Journal of biological chemistry.

[48]  D. Baltimore,et al.  I kappa B: a specific inhibitor of the NF-kappa B transcription factor. , 1988, Science.

[49]  K. Nakai,et al.  PSORT: a program for detecting sorting signals in proteins and predicting their subcellular localization. , 1999, Trends in biochemical sciences.