Identification of ku and dna-pkcs domains responsible for ku heterodimer and dna-pk holoenzyme assembly

Robert B. Cary, Fanqing Chen, Zhiyuan Shen, Akihiro Kurimasa, David J. Chen

Research output: Contribution to journalArticle

Abstract

DNA dependent protein kinase (DNA-PK) may play a critical role in DNA double strand break repair in mammalian cells. The DNA-PK holoenzyme is composed of the Ku heterodimer and a large catalytic subunit (DNA-PKcs). Although current understanding of the structure of DNA-PK is limited, it is thought that the Ku heterodimer is responsible for targeting the catalytic subunit to DNA ends. To better understand the structure of DNA-PK we are using a combination of in vivo and in vitro approaches to map the regions of Ku and the DNA-PK catalytic subunit responsible for holoenzyme assembly. The yeast two-hybrid system provides a powerful means of identifying domains responsible for mediating protein-protein interactions. We have identified a region of Ku80, outside of and C-terminal to the putative leucine zipper, that is capable of mediating interaction with Ku70 in the yeast two-hybrid system. Currently, in vitro studies are being used to confirm the ability of this region to interact with Ku70. Analysis of DNA-PKcs fragments in the yeast two- hybrid system are underway to identify regions of the catalytic subunit responsible for interaction with Ku. The association of DNA-PK with DNA is being examined by a combination of election and atomic force microscopy.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number6
StatePublished - 1996

Fingerprint

DNA-Activated Protein Kinase
Holoenzymes
Two-Hybrid System Techniques
Catalytic Domain
DNA
protein kinases
Hybrid systems
Yeast
protein subunits
two hybrid system techniques
Catalytic DNA
Leucine Zippers
Double-Stranded DNA Breaks
Atomic Force Microscopy
yeasts
Ku Autoantigen
Atomic force microscopy
Proteins
Repair
Cells

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Cary, R. B., Chen, F., Shen, Z., Kurimasa, A., & Chen, D. J. (1996). Identification of ku and dna-pkcs domains responsible for ku heterodimer and dna-pk holoenzyme assembly. FASEB Journal, 10(6).

Identification of ku and dna-pkcs domains responsible for ku heterodimer and dna-pk holoenzyme assembly. / Cary, Robert B.; Chen, Fanqing; Shen, Zhiyuan; Kurimasa, Akihiro; Chen, David J.

In: FASEB Journal, Vol. 10, No. 6, 1996.

Research output: Contribution to journalArticle

Cary, RB, Chen, F, Shen, Z, Kurimasa, A & Chen, DJ 1996, 'Identification of ku and dna-pkcs domains responsible for ku heterodimer and dna-pk holoenzyme assembly', FASEB Journal, vol. 10, no. 6.
Cary, Robert B. ; Chen, Fanqing ; Shen, Zhiyuan ; Kurimasa, Akihiro ; Chen, David J. / Identification of ku and dna-pkcs domains responsible for ku heterodimer and dna-pk holoenzyme assembly. In: FASEB Journal. 1996 ; Vol. 10, No. 6.
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