ABSTRACT
The heterodimeric Ku protein (composed of the Ku 86 and Ku 70 sub-units) is a nuclear protein which binds to DNA termini without sequence specificity. Ku is the DNA-targeting component of the large catalytic sub-unit of the DNA-dependent protein kinase complex that is required for the repair of DNA double-strand breaks in mammalian cells. We studied the expression and function of Ku/DNA-PK during granulocytic differentiation of two human promyelocytic cell lines, HL60 and NB4, a process associated to decreased radiation resistance. After 3 days exposure to differentiating agents (either all-trans-retinoic acid or DMSO), Ku binding to double stranded (ds)-DNA ends declined dramatically whereas Ku protein levels remain unchanged. The nuclear, but not cytoplasmic, fraction of differentiated HL60 cells extracts exhibited a heat-sensitive inhibitory activity towards DNA binding of recombinant Ku heterodimer. We further demonstrate that immunoprecipitation of Ku is impaired in extracts from differentiated cells by using two antibodies that recognize epitopes within the C-terminus DNA binding domains of Ku 70 and Ku 86 proteins. These results favor the hypothesis of a protein interacting with Ku that would prevent DNA binding of heterodimerized Ku protein by steric hindrance.
Subject(s)
Antigens, Nuclear , DNA Helicases , DNA-Binding Proteins/metabolism , Granulocytes/physiology , Myeloid Progenitor Cells/physiology , Nuclear Proteins/metabolism , Antibodies, Monoclonal/immunology , Cell Differentiation , Cell Extracts/analysis , Cell Line , Cell Nucleus/metabolism , DNA Repair , DNA-Binding Proteins/immunology , DNA-Binding Proteins/physiology , Dimerization , Down-Regulation , Epitopes/immunology , HL-60 Cells , Humans , Ku Autoantigen , Nuclear Proteins/immunology , Nuclear Proteins/physiology , Precipitin Tests , Protein Structure, TertiaryABSTRACT
Direct measurements of total reaction cross sections (sigma R) have been performed in the energy range of 10-300 MeV/nucleon for heavy ion collisions. A decrease of sigma R with increasing energy was observed for a wide range of masses of the colliding systems. The data suggest that sigma R reaches a minimum located around 300 MeV/nucleon independently of the projectile target combination. A dependence of sigma R on mass asymmetry of the svstem is also demonstrated. Trends of sigma R in this energy range are well reproduced by the predictions of a simple microscopic model based on individual nucleon-nucleon collisions. Our data have been employed in this framework to derive a new semi-empirical parametrization of sigma R. Most of the experimental results in the intermediate and high energy range have been reproduced by this parametrization using a single energy-dependent parameter.
Subject(s)
Elementary Particle Interactions , Heavy Ions , Models, Theoretical , Nuclear Physics , Carbon , Elements , Energy Transfer , Iron Isotopes , Neon , Neutrons , Oxygen , Particle Accelerators , Scattering, Radiation , Zinc IsotopesABSTRACT
Experimental results are presented from vacuum-ultraviolet free-electron laser (FEL) operating in the self-amplified spontaneous emission (SASE) mode. The generation of ultrashort radiation pulses became possible due to specific tailoring of the bunch charge distribution. A complete characterization of the linear and nonlinear modes of the SASE FEL operation was performed. At saturation the FEL produces ultrashort pulses (30-100 fs FWHM) with a peak radiation power in the GW level and with full transverse coherence. The wavelength was tuned in the range of 95-105 nm.