Function domains of Ku proteins as molecular targets

Summary

Principal Investigator: Fanqing Chen
Affiliation: Lawrence Berkeley National Laboratory
Country: USA
Abstract: Radiation is one of the most efficient therapeutic agents for certain types of cancer. However, cytotoxicity induced by ionizing radiation limits the radiation dosage that can be administrated to patients. Alternatively, several DNA modification agents and other adjunct treatments have been utilized as "radiosensitizers" to combine with the standard radiotherapy, thus enhancing the radiosensitivity of the targeted cancer cells. Because of the inherent nonspecificity of current radiosensitizers, the enhancement ratio is relatively moderate, and the applicable range clinically, is limited by side effects. Meanwhile, it has become clear that disruption of DNA repair might dramatically increase radiosensitivity. In this research, we propose to explore the possibility of using DNA double-strand break repair proteins as novel molecular targets for the development of radiosensitizers. Specifically, we will focus on the Ku70 and Ku80 proteins (Ku70/80 heterodimers), which are critical components in the repair of ionizing radiation-induced double-strand breaks in mammalian cells. We plan to identify and validate specific domains of the Ku70/80 heterodimer as novel protein targets for identifying effective radiosensitizers. This study proposes to dissect Ku functional domain using genetic, biochemical, proteolytic digestion/mass spectrometry data, and to express the domains in soluble protein form. Using the expressed recombinant protein domains, we will demonstrate the validity of Ku domains as a molecular target with in vitro biochemical assays, and in vivo dominant negative phenotyping. We also intend to use the combinatorial phage display technique to screen for high-affinity peptides for the interested domains. In viva inhibition studies will be performed by introducing the peptide into cancer cells in a retroviral vector. We will also analyze the structural relationship between the peptide inhibitors and their affected functional domains by NMR. The ultimate goal for this study is to identify the Ku domains that can be used as molecular targets for inhibitory radiosensitizers, and to generate more specific inhibitors/effectors that direct more distinct damages to the cancer cells under inadiation.
Funding Period: 2002-04-26 - 2006-03-31
more information: NIH RePORT

Top Publications

  1. pmc Sphingolipids are involved in N-methyl-N'-nitro-N-nitrosoguanidine-induced epidermal growth factor receptor clustering
    Yun Huang
    Department of Pathology and Pathophysiology, Center for Environmental Genomics, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310031, China
    Biochem Biophys Res Commun 330:430-8. 2005
  2. doi A nanoplasmonic molecular ruler for measuring nuclease activity and DNA footprinting
    Gang L Liu
    Nat Nanotechnol 1:47-52. 2006
  3. ncbi Peptide-nanoparticle hybrid SERS probe for dynamic detection of active cancer biomarker enzymes
    Gang L Liu
    Department of Bioengineering, University of California at Berkeley, CA 94720, USA
    Conf Proc IEEE Eng Med Biol Soc 1:795-8. 2006
  4. ncbi Peptide-nanoparticle hybrid SERS probes for optical detection of protease activity
    Gang L Liu
    Department of Bioengineering, Biomolecular Nanotechnology Center, Berkeley Sensor and Actuator Center, University of California, Berkeley, CA 94720, USA
    J Nanosci Nanotechnol 7:2323-30. 2007
  5. ncbi Charge-associated effects of fullerene derivatives on microbial structural integrity and central metabolism
    Yinjie J Tang
    Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA
    Nano Lett 7:754-60. 2007
  6. ncbi A novel type of PTD, common helix-loop-helix motif, could efficiently mediate protein transduction into mammalian cells
    Jing Chen
    State Key Laboratory of Genetic Engineering, School of Life Science, University of Fudan, Shanghai 200433, China
    Biochem Biophys Res Commun 347:931-40. 2006
  7. pmc A comparative study of using comet assay and gammaH2AX foci formation in the detection of N-methyl-N'-nitro-N-nitrosoguanidine-induced DNA damage
    Yanke Yu
    Department of Public Health, Center for Environmental Genomics, Zhejiang University School of Medicine, 353 Yan An Road, Hangzhou, Zhejiang 310031, China
    Toxicol In Vitro 20:959-65. 2006
  8. pmc Lithographic techniques and surface chemistries for the fabrication of PEG-passivated protein microarrays
    Balaji Kannan
    Department of Mechanical Engineering, University of California, Berkeley, CA 94720, USA
    Biosens Bioelectron 21:1960-7. 2006
  9. pmc DNA damage evaluated by gammaH2AX foci formation by a selective group of chemical/physical stressors
    Chunxian Zhou
    Department of Public Health, Institute for Toxicology, Zhejiang University School of Medicine, 353 Yan An Road, Hangzhou, Zhejiang 310031, China
    Mutat Res 604:8-18. 2006
  10. pmc Molecular characterization of the cytotoxic mechanism of multiwall carbon nanotubes and nano-onions on human skin fibroblast
    Lianghao Ding
    Life Sciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720, USA
    Nano Lett 5:2448-64. 2005

Scientific Experts

  • Gang L Liu
  • Balaji Kannan
  • Jun Yang
  • Fanqing F Chen
  • Yun Huang
  • Fanqing Frank Chen
  • Yanke Yu
  • Chunxian Zhou
  • Cheng Sun
  • Omeed Elboudwarej
  • Yinjie J Tang
  • Jing Chen
  • Wen Zhu
  • Huiling Diao
  • Jing Shen
  • Lianghao Ding
  • Charles S Craik
  • Yazmin T Rosa-Bauza
  • Kai Hung Su
  • Marc A Shuman
  • Jason Valentine
  • Jonathan A Ellman
  • Xiang Zhang
  • Bipasha Mukherjee
  • Chul Hyun Kim
  • Jared M Ashcroft
  • Jay D Keasling
  • Carolyn Larabell
  • Ding Chen
  • Bipasha Murkhejee
  • Guangwei Min
  • Jun Lu
  • Jiaxin Zhang
  • Zhongxiang Li
  • Huiling Wu
  • Yin Huang
  • Yayun Dai
  • Daru Lu
  • Lei Chen
  • Ge Li
  • John P Selegue
  • Tingting Zhang
  • Joe W Gray
  • Ting Wang
  • Yan ke Yu
  • Patrick A Cooke
  • Huijian Jiang
  • Jackie Stilwell
  • Yingnian Yu

Detail Information

Publications12

  1. pmc Sphingolipids are involved in N-methyl-N'-nitro-N-nitrosoguanidine-induced epidermal growth factor receptor clustering
    Yun Huang
    Department of Pathology and Pathophysiology, Center for Environmental Genomics, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310031, China
    Biochem Biophys Res Commun 330:430-8. 2005
    ..Taken together, these data suggested that sphingolipids are involved in MNNG-induced receptor clustering; however, the specific species involved may be different from those involved in EGF-mediated receptor clustering...
  2. doi A nanoplasmonic molecular ruler for measuring nuclease activity and DNA footprinting
    Gang L Liu
    Nat Nanotechnol 1:47-52. 2006
  3. ncbi Peptide-nanoparticle hybrid SERS probe for dynamic detection of active cancer biomarker enzymes
    Gang L Liu
    Department of Bioengineering, University of California at Berkeley, CA 94720, USA
    Conf Proc IEEE Eng Med Biol Soc 1:795-8. 2006
    ..Peptide-conjugated nanocrescents should also be applicable for measuring the intercellular and intracellular activity of other cancer-related proteases and protease activity profiling-enabled cancer cell identification...
  4. ncbi Peptide-nanoparticle hybrid SERS probes for optical detection of protease activity
    Gang L Liu
    Department of Bioengineering, Biomolecular Nanotechnology Center, Berkeley Sensor and Actuator Center, University of California, Berkeley, CA 94720, USA
    J Nanosci Nanotechnol 7:2323-30. 2007
    ....
  5. ncbi Charge-associated effects of fullerene derivatives on microbial structural integrity and central metabolism
    Yinjie J Tang
    Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA
    Nano Lett 7:754-60. 2007
    ..The results presented here favor the hypothesis that fullerenes cause more membrane stress 4-6 than perturbation to energy metabolism.7...
  6. ncbi A novel type of PTD, common helix-loop-helix motif, could efficiently mediate protein transduction into mammalian cells
    Jing Chen
    State Key Laboratory of Genetic Engineering, School of Life Science, University of Fudan, Shanghai 200433, China
    Biochem Biophys Res Commun 347:931-40. 2006
    ....
  7. pmc A comparative study of using comet assay and gammaH2AX foci formation in the detection of N-methyl-N'-nitro-N-nitrosoguanidine-induced DNA damage
    Yanke Yu
    Department of Public Health, Center for Environmental Genomics, Zhejiang University School of Medicine, 353 Yan An Road, Hangzhou, Zhejiang 310031, China
    Toxicol In Vitro 20:959-65. 2006
    ..Therefore, gammaH2AX may be used as a sensitive indicator for DNA damage...
  8. pmc Lithographic techniques and surface chemistries for the fabrication of PEG-passivated protein microarrays
    Balaji Kannan
    Department of Mechanical Engineering, University of California, Berkeley, CA 94720, USA
    Biosens Bioelectron 21:1960-7. 2006
    ..The effectiveness of this microarray technology is demonstrated by patterning fluorescent streptavidin and a fluorescent secondary antibody using the well-known and highly specific interaction between biotin and streptavidin...
  9. pmc DNA damage evaluated by gammaH2AX foci formation by a selective group of chemical/physical stressors
    Chunxian Zhou
    Department of Public Health, Institute for Toxicology, Zhejiang University School of Medicine, 353 Yan An Road, Hangzhou, Zhejiang 310031, China
    Mutat Res 604:8-18. 2006
    ....
  10. pmc Molecular characterization of the cytotoxic mechanism of multiwall carbon nanotubes and nano-onions on human skin fibroblast
    Lianghao Ding
    Life Sciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720, USA
    Nano Lett 5:2448-64. 2005
    ....
  11. ncbi A lipidomic study of the effects of N-methyl-N'-nitro-N-nitrosoguanidine on sphingomyelin metabolism
    Yun Huang
    Department of Pathology and Pathophysiology, Center for Environmental Genomics, Zhejiang University School of Medicine, Hangzhou 310031, China
    Acta Biochim Biophys Sin (Shanghai) 37:515-24. 2005
    ..It also prevented some of the effects of MNNG, as well as the redistribution of ASM. Taken together, these data suggested that the lipidomic approach is highly effective for the systematic analysis of cellular lipids metabolism...
  12. pmc Time-resolved single-step protease activity quantification using nanoplasmonic resonator sensors
    Cheng Sun
    Nanoscale Science and Engineering Center, University of California, Berkeley, CA 94720, USA
    ACS Nano 4:978-84. 2010
    ..This method offers a fast, sensitive, accurate, and one-step approach to detect the proteases' activities in very small sample volumes...