Dimerization, translocation and localization of Ku70 and Ku80 proteins.

The Ku protein is a complex of two subunits, Ku70 and Ku80, and was originally identified as an autoantigen recognized by the sera of patients with autoimmune diseases. The Ku protein plays a key role in multiple nuclear processes, e.g., DNA repair, chromosome maintenance, transcription regulation, and V(D)J recombination. The mechanism underlying the regulation of all the diverse functions of Ku is still unclear, although it seems that Ku is a multifunctional protein that works in nuclei. On the other hand, several studies have reported cytoplasmic or cell surface localization of Ku in various cell types. To clarify the fundamental characteristics of Ku, we have examined the expression, heterodimerization, subcellular localization, chromosome location, and molecular mechanisms of the nuclear transport of Ku70 and Ku80. The mechanism that regulates for nuclear localization of Ku70 and Ku80 appears to play, at least in part, a key role in regulating the physiological function of Ku in vivo.

[1]  F. Alt,et al.  Ku80: product of the XRCC5 gene and its role in DNA repair and V(D)J recombination. , 1994, Science.

[2]  S. Lees-Miller The DNA-dependent protein kinase, DNA-PK: 10 years and no ends in sight. , 1996, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[3]  K. Khanna,et al.  DNA double-strand breaks: signaling, repair and the cancer connection , 2001, Nature Genetics.

[4]  R. Geha,et al.  Ku in the cytoplasm associates with CD40 in human B cells and translocates into the nucleus following incubation with IL-4 and anti-CD40 mAb. , 1999, Immunity.

[5]  Thomas Ried,et al.  DNA repair protein Ku80 suppresses chromosomal aberrations and malignant transformation , 2000, Nature.

[6]  P. Jeggo,et al.  DNA double-strand break repair and V(D)J recombination: involvement of DNA-PK. , 1995, Trends in biochemical sciences.

[7]  K. Muegge,et al.  Defining the Minimal Domain of Ku80 for Interaction with Ku70* , 1997, The Journal of Biological Chemistry.

[8]  Yunmei Ma,et al.  Hairpin Opening and Overhang Processing by an Artemis/DNA-Dependent Protein Kinase Complex in Nonhomologous End Joining and V(D)J Recombination , 2002, Cell.

[9]  J. Cigudosa,et al.  Mammalian Ku86 mediates chromosomal fusions and apoptosis caused by critically short telomeres , 2002, The EMBO journal.

[10]  W. Dynan,et al.  Interaction of Ku protein and DNA-dependent protein kinase catalytic subunit with nucleic acids. , 1998, Nucleic acids research.

[11]  H. Hameister,et al.  MCM4 and PRKDC, human genes encoding proteins MCM4 and DNA-PKcs, are close neighbours located on chromosome 8q12-->q13. , 1997, Cytogenetics and cell genetics.

[12]  M. Koike,et al.  Ku70 can translocate to the nucleus independent of Ku80 translocation and DNA-PK autophosphorylation. , 2000, Biochemical and biophysical research communications.

[13]  Jeremy M. Stark,et al.  Double-strand breaks and tumorigenesis. , 2001, Trends in cell biology.

[14]  M. Koike,et al.  The nuclear localization signal of the human Ku70 is a variant bipartite type recognized by the two components of nuclear pore-targeting complex. , 1999, Experimental cell research.

[15]  T. de Lange,et al.  Ku Binds Telomeric DNA in Vitro * , 1999, The Journal of Biological Chemistry.

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

[17]  Michel C. Nussenzweig,et al.  Genomic Instability in Mice Lacking Histone H2AX , 2002, Science.

[18]  T. Mimori,et al.  Chromosomal localization of the mouse and rat DNA double-strand break repair genes Ku p70 and Ku p80/XRCC5 and their mRNA expression in various mouse tissues. , 1996, Genomics.

[19]  N. Imamoto,et al.  Differential Modes of Nuclear Localization Signal (NLS) Recognition by Three Distinct Classes of NLS Receptors* , 1997, The Journal of Biological Chemistry.

[20]  Manabu Koike,et al.  Ku80 can translocate to the nucleus independent of the translocation of Ku70 using its own nuclear localization signal , 1999, Oncogene.

[21]  T. Kuramoto,et al.  A rat genetic linkage map and comparative maps for mouse or human homologous rat genes , 1994, Mammalian Genome.

[22]  Y. Hurd,et al.  Autoantigen Ku in the brain. Developmentally regulated expression and subcellular localization , 1998, Neuroreport.

[23]  J. Hoeijmakers,et al.  Chromosomal stability and the DNA double-stranded break connection , 2001, Nature Reviews Genetics.

[24]  M. Nussenzweig,et al.  Requirement for Ku80 in growth and immunoglobulin V(D)J recombination , 1996, Nature.

[25]  M. Lieber,et al.  Protein-protein and protein-DNA interaction regions within the DNA end-binding protein Ku70-Ku86 , 1996, Molecular and cellular biology.

[26]  B. Nelms,et al.  hMre11 and hRad50 nuclear foci are induced during the normal cellular response to DNA double-strand breaks , 1997, Molecular and cellular biology.

[27]  J. Quinn,et al.  The nuclear autoimmune antigen Ku is also present on the cell surface. , 1992, Autoimmunity.

[28]  E. Blackburn,et al.  Ku is associated with the telomere in mammals. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[29]  P. Jeggo,et al.  Molecular and biochemical characterization of xrs mutants defective in Ku80 , 1997, Molecular and cellular biology.

[30]  T. Mimori,et al.  Subcellular localization and protein-protein interaction regions of Ku proteins. , 1998, Biochemical and biophysical research communications.

[31]  John A Tainer,et al.  DNA double-strand break repair from head to tail. , 2002, Current opinion in structural biology.

[32]  A. Kuroiwa,et al.  Cytogenetic mapping of 31 functional genes on chicken chromosomes by direct R-banding FISH , 1999, Cytogenetic and Genome Research.

[33]  J. Blanchard,et al.  Chromosomal location and expression of the genes coding for Ku p70 and p80 in human cell lines and normal tissues. , 1994, Cytogenetics and cell genetics.

[34]  Y. Matsuda,et al.  Expression and chromosome location of hamster Ku70 and Ku80 , 2001, Cytogenetic and Genome Research.

[35]  T. Morita,et al.  Immunolocalization of Ku-proteins (p80/p70): localization of p70 to nucleoli and periphery of both interphase nuclei and metaphase chromosomes. , 1992, Experimental cell research.

[36]  M. Hayashi,et al.  Deficiency in Nuclear Accumulation of G22p1 and Xrcc5 Proteins in Hyper-radiosensitive Long-Evans Cinnamon (LEC) Rat Cells after X Irradiation , 2002, Radiation research.

[37]  F. Alt,et al.  Growth retardation and leaky SCID phenotype of Ku70-deficient mice. , 1997, Immunity.

[38]  M. Lieber,et al.  Interaction of DNA‐dependent protein kinase with DNA and with Ku: biochemical and atomic‐force microscopy studies , 1997, The EMBO journal.

[39]  C. Ling,et al.  Ku70: a candidate tumor suppressor gene for murine T cell lymphoma. , 1998, Molecular cell.

[40]  R. Ochs,et al.  Purification of an 86-70 kDa nuclear DNA-associated protein complex. , 1985, Biochimica et biophysica acta.

[41]  D. Ward,et al.  Gene for the catalytic subunit of the human DNA-activated protein kinase maps to the site of the XRCC7 gene on chromosome 8. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[42]  R. Haché,et al.  Nuclear localization of Ku antigen is promoted independently by basic motifs in the Ku70 and Ku80 subunits. , 2001, Journal of cell science.

[43]  M. Connelly,et al.  The promoters for human DNA-PKcs (PRKDC) and MCM4: divergently transcribed genes located at chromosome 8 band q11. , 1998, Genomics.

[44]  A. Fischer,et al.  Artemis, a Novel DNA Double-Strand Break Repair/V(D)J Recombination Protein, Is Mutated in Human Severe Combined Immune Deficiency , 2001, Cell.

[45]  G. Tsujimoto,et al.  Differential subcellular localization of DNA-dependent protein kinase components Ku and DNA-PKcs during mitosis. , 1999, Journal of cell science.

[46]  Shengfang Jin,et al.  Double‐strand break repair by Ku70 requires heterodimerization with Ku80 and DNA binding functions , 1997, The EMBO journal.

[47]  J. Walker,et al.  Structure of the Ku heterodimer bound to DNA and its implications for double-strand break repair , 2001, Nature.

[48]  D. Ward,et al.  Nuclear foci of mammalian Rad51 recombination protein in somatic cells after DNA damage and its localization in synaptonemal complexes. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[49]  S. Jackson,et al.  Identification of bacterial homologues of the Ku DNA repair proteins , 2001, FEBS letters.

[50]  M. Hande,et al.  Ku acts in a unique way at the mammalian telomere to prevent end joining. , 2000, Genes & development.

[51]  S. Pongor,et al.  A somatostatin analogue induces translocation of Ku 86 autoantigen from the cytosol to the nucleus in colon tumour cells. , 1998, Cellular signalling.

[52]  G. Chu,et al.  Ku86 defines the genetic defect and restores X-ray resistance and V(D)J recombination to complementation group 5 hamster cell mutants , 1996, Molecular and cellular biology.

[53]  Reeves Wh Antinuclear antibodies as probes to explore the structural organization of the genome. , 1987 .

[54]  D. Weaver,et al.  Regulation and repair of double-strand DNA breaks. , 1996, Critical reviews in eukaryotic gene expression.

[55]  P. Jeggo Identification of genes involved in repair of DNA double-strand breaks in mammalian cells. , 1998, Radiation research.

[56]  O. Hammarsten,et al.  DNA-dependent protein kinase: DNA binding and activation in the absence of Ku. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[57]  S. Jackson,et al.  Mammalian DNA double-strand break repair protein XRCC4 interacts with DNA ligase IV , 1997, Current Biology.

[58]  Molly A Bogue,et al.  Ku86-Deficient Mice Exhibit Severe Combined Immunodeficiency and Defective Processing of V(D)J Recombination Intermediates , 1996, Cell.

[59]  Hans H. Cheng,et al.  A consensus linkage map of the chicken genome. , 2000, Genome research.

[60]  F. Chen,et al.  A central region of Ku80 mediates interaction with Ku70 in vivo. , 1998, Nucleic acids research.

[61]  S. Jackson,et al.  Mapping of protein-protein interactions within the DNA-dependent protein kinase complex. , 1999, Nucleic acids research.

[62]  R. Leber,et al.  The XRCC4 Gene Product Is a Target for and Interacts with the DNA-dependent Protein Kinase* , 1998, The Journal of Biological Chemistry.

[63]  M. Blasco,et al.  Mammalian Ku86 protein prevents telomeric fusions independently of the length of TTAGGG repeats and the G‐strand overhang , 2000, EMBO reports.

[64]  Y. Yoneda How proteins are transported from cytoplasm to the nucleus. , 1997, Journal of biochemistry.

[65]  S. Jackson,et al.  The DNA-dependent protein kinase , 1999 .

[66]  M. Hayashi,et al.  Hypertonic Treatment Inhibits Radiation-Induced Nuclear Translocation of the Ku Proteins G22p1 (Ku70) and Xrcc5 (Ku80) in Rat Fibroblasts , 2001, Radiation research.

[67]  S. Jackson,et al.  Ku, a DNA repair protein with multiple cellular functions? , 1999, Mutation research.

[68]  Y. Matsuda,et al.  Construction of comparative cytogenetic maps of the Chinese hamster to mouse, rat and human , 2004, Chromosome Research.

[69]  H. Vogel,et al.  Analysis of ku80-Mutant Mice and Cells with Deficient Levels of p53 , 2000, Molecular and Cellular Biology.

[70]  M. Lagally,et al.  In situ visualization of DNA double-strand break repair in human fibroblasts. , 1998, Science.

[71]  J. Steitz,et al.  Characterization of the DNA-binding protein antigen Ku recognized by autoantibodies from patients with rheumatic disorders. , 1986, The Journal of biological chemistry.

[72]  R. West,et al.  Productive and Nonproductive Complexes of Ku and DNA-Dependent Protein Kinase at DNA Termini , 1998, Molecular and Cellular Biology.

[73]  H. Taylor,et al.  HOXA10 is expressed in response to sex steroids at the time of implantation in the human endometrium. , 1998, The Journal of clinical investigation.

[74]  B. Lehnert,et al.  DNA double-strand break repair proteins are required to cap the ends of mammalian chromosomes. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[75]  Reeves Wh,et al.  Antibodies to the p70/p80 (Ku) antigens in systemic lupus erythematosus. , 1992 .

[76]  J. Petrini,et al.  DNA Damage-Dependent Nuclear Dynamics of the Mre11 Complex , 2001, Molecular and Cellular Biology.

[77]  E. Kuff,et al.  Intracellular redistribution of Ku immunoreactivity in response to cell-cell contact and growth modulating components in the medium. , 1996, Journal of cell science.

[78]  J. Blanchard,et al.  Hypersensitivity of Ku-Deficient Cells toward the DNA Topoisomerase II Inhibitor ICRF-193 Suggests a Novel Role for Ku Antigen during the G2 and M Phases of the Cell Cycle , 1998, Molecular and Cellular Biology.

[79]  M. Blasco,et al.  The Absence of the DNA-Dependent Protein Kinase Catalytic Subunit in Mice Results in Anaphase Bridges and in Increased Telomeric Fusions with Normal Telomere Length and G-Strand Overhang , 2001, Molecular and Cellular Biology.

[80]  M. Koike,et al.  Dimerization and Nuclear Localization of Ku Proteins* , 2001, The Journal of Biological Chemistry.

[81]  B. Prabhakar,et al.  Cell surface expression of the 70-kD component of Ku, a DNA-binding nuclear autoantigen. , 1990, The Journal of clinical investigation.

[82]  L. L. Li,et al.  Cell cycle-dependent migration of the DNA-binding protein Ku80 into nucleoli. , 1992, Experimental cell research.

[83]  S. Jhiang,et al.  Expression of the Ku protein during cell proliferation. , 1991, Biochimica et biophysica acta.

[84]  T. Mimori,et al.  Characterization of a high molecular weight acidic nuclear protein recognized by autoantibodies in sera from patients with polymyositis-scleroderma overlap. , 1981, The Journal of clinical investigation.

[85]  K. Anderson,et al.  The 86-kD subunit of Ku autoantigen mediates homotypic and heterotypic adhesion of multiple myeloma cells. , 1998, The Journal of clinical investigation.

[86]  P. Jeggo DNA-PK: at the cross-roads of biochemistry and genetics. , 1997, Mutation research.

[87]  C. Anderson,et al.  DNA damage and the DNA-activated protein kinase. , 1993, Trends in biochemical sciences.

[88]  W. Rodgers,et al.  Transient Association of Ku with Nuclear Substrates Characterized Using Fluorescence Photobleaching1 , 2002, The Journal of Immunology.

[89]  K. Myung,et al.  Identification of Two Domains of the p70 Ku Protein Mediating Dimerization with p80 and DNA Binding* , 1998, The Journal of Biological Chemistry.

[90]  R. Jessberger,et al.  Expression of DNA-dependent protein kinase holoenzyme upon induction of lymphocyte differentiation and V(D)J recombination. , 1996, European journal of biochemistry.

[91]  M. Hande,et al.  Effects of DNA nonhomologous end-joining factors on telomere length and chromosomal stability in mammalian cells , 2001, Current Biology.

[92]  Fengfei Wang,et al.  Translocation of Ku86/Ku70 to the multiple myeloma cell membrane: functional implications. , 2002, Experimental hematology.

[93]  F. Alt,et al.  A targeted DNA-PKcs-null mutation reveals DNA-PK-independent functions for KU in V(D)J recombination. , 1998, Immunity.