Cell Signaling Technology

Product Pathways - Protein Stability

DDB-1 Antibody #5428

Applications Reactivity Sensitivity MW (kDa) Source
W H M R Mk Endogenous 127 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

DDB-1 Antibody detects endogenous levels of total DDB-1 protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human DDB-1 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using DDB-1 Antibody.

Background

Damaged DNA-Binding Protein (DDB) consists of a 127 kDa subunit (DDB-1) and a 48 kDa subunit (DDB-2) that contribute to the formation of the UV-damaged DNA-binding protein complex (UV-DDB) (1-3). In conjunction with CUL4A and ROC-1, the UV-DDB complex forms an E3 ubiquitin ligase that recognizes a broad spectrum of DNA lesions such as cyclobutane pyrimidine dimers, 6-4 photoproducts, apurinic sites and short mismatches. The complex polyubiquitinates components of the nucleotide excision repair pathway (4-6). Loss of DDB activity has been identified in a subset of xeroderma pigmentosum complementation group E (XP-E) patients and has been linked to the deficient repair of cyclobutane pyrimidine dimers in cells derived from these patients (7-10).

DDB-1 is a relatively abundant protein that is vital for normal cell function and is evolutionarily conserved in mammals, insects, worms and plants. Unlike DDB-2, lesions in DDB-1 have yet to be indentified in XP-E patients. In association with ROC-1 and CUL4A, DDB-1 functions to recruit substrate-specific targeting subunits, generally known as DCAFs or CDWs, to CUL4-RING E3 ubiquitin-protein ligase complexes (11,12). Ubiquitination of histone H2A, histone H3 and histone H4 at sites of UV-induced DNA damage by the DDB1-DDB2-CUL4A-ROC1 E3 ubiquitin-protein ligase complex may facilitate their removal from the nucleosome in order to promote DNA repair (13-15). DDB-1, in association with other CUL4-based E3 ligase complexes, has also been found to be a regulator of mTOR signaling (16,17).

  1. Reardon, J.T. et al. (1993) J Biol Chem 268, 21301-8.
  2. Keeney, S. et al. (1993) J Biol Chem 268, 21293-300.
  3. Hwang, B.J. and Chu, G. (1993) Biochemistry 32, 1657-66.
  4. Chu, G. and Chang, E. (1990) Proc Natl Acad Sci USA 87, 3324-7.
  5. Hirschfeld, S. et al. (1990) Mol Cell Biol 10, 2041-8.
  6. Payne, A. and Chu, G. (1994) Mutat Res 310, 89-102.
  7. Chu, G. and Chang, E. (1988) Science 242, 564-7.
  8. Nichols, A.F. et al. (1996) J Biol Chem 271, 24317-20.
  9. Kataoka, H. and Fujiwara, Y. (1991) Biochem Biophys Res Commun 175, 1139-43.
  10. Keeney, S. et al. (1992) Mutat Res 273, 49-56.
  11. He, Y.J. et al. (2006) Genes Dev 20, 2949-54.
  12. Lee, J. and Zhou, P. (2007) Mol Cell 26, 775-80.
  13. Wang, H. et al. (2006) Mol Cell 22, 383-94.
  14. Kapetanaki, M.G. et al. (2006) Proc Natl Acad Sci USA 103, 2588-93.
  15. Guerrero-Santoro, J. et al. (2008) Cancer Res 68, 5014-22.
  16. Ghosh, P. et al. (2008) Cell Cycle 7, 373-81.
  17. Hu, J. et al. (2008) Genes Dev 22, 866-71.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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