Ysis of key elements of identified DDB2 protein sub-complexes. DDB1 and Cullin4A of the ubiquitin Dimethylenastron supplier ligase sub-complex at the same time as CSN5 with the COP9 signalosome subcomplex are detected along with FLAG-DDB2. Acknowledgments We thank members from the Iglehart laboratory, and Dr. Sylvie Vincent for valuable discussions. Author Contributions Conceived and made the experiments: MD JDI JBL. Performed the experiments: MD ASC JG DJE MRS GLF. Analyzed the information: MD 1676428 JBL ASC JG DS. Contributed reagents/materials/analysis tools: SI. Wrote the paper: MD JDI JBL DEF SI DS. References 1. Jackson SP, K162 Bartek J The DNA-damage response in human biology and disease. Nature 461: 10711078. two. Keeney S, Chang GJ, Linn S Characterization of a human DNA damage binding protein implicated in xeroderma pigmentosum E. J Biol Chem 268: 2129321300. three. Takao M, Abramic M, Moos M Jr, Otrin VR, Wootton JC, et al. A 127 kDa element of a UV-damaged DNA-binding complicated, which can be defective in some xeroderma pigmentosum group E individuals, is homologous to a slime mold protein. Nucleic Acids Res 21: 41114118. four. Vermeulen W Dynamics of mammalian NER proteins. DNA Repair 10: 760771. five. Curtin NJ DNA repair dysregulation from cancer driver to therapeutic target. Nat Rev Cancer 12: 801817. six. Ravanat JL, Douki T, Cadet J Direct and indirect effects of UV radiation on DNA and its elements. J Photochem Photobiol B 63: 88102. 7. Cadet J, Sage E, Douki T Ultraviolet radiation-mediated damage to cellular DNA. Mutat Res 571: 317. 8. Tang JY, Hwang BJ, Ford JM, Hanawalt Computer, Chu G Xeroderma pigmentosum p48 gene enhances worldwide genomic repair and suppresses UVinduced mutagenesis. Mol Cell 5: 737744. 9. Chu G, Chang E Xeroderma pigmentosum group E cells lack a nuclear element that binds to damaged DNA. Science 242: 564567. ten. Alekseev S, Kool H, Rebel H, Fousteri M, Moser J, et al. Enhanced DDB2 expression protects mice from carcinogenic effects of chronic UV-B irradiation. Cancer Res 65: 1029810306. 11. Moser J, Volker M, Kool H, Alekseev S, Vrieling H, et al. The UVdamaged DNA binding protein mediates effective targeting of the nucleotide 12. excision repair complex to UV-induced photo lesions. DNA Repair 4: 571582. Fujiwara Y, Masutani C, Mizukoshi T, Kondo J, Hanaoka F, et al. Characterization of DNA recognition by the human UV-damaged DNAbinding protein. J Biol Chem 274: 2002720033. Reardon JT, Nichols AF, Keeney S, Smith CA, Taylor JS, et al. Comparative evaluation of binding of human broken DNA-binding protein and Escherichia coli harm recognition protein to the key ultraviolet photoproducts: TT, TT, TT, and TT. J Biol Chem 268: 2130121308. Treiber DK, Chen Z, Essigmann JM An ultraviolet light-damaged DNA recognition protein absent in xeroderma pigmentosum group E cells binds selectively to pyrimidine pyrimidone photoproducts. Nucleic Acids Res 20: 58055810. Fischer ES, Scrima A, Bohm K, Matsumoto S, Lingaraju GM, et al. The molecular basis of CRL4DDB2/CSA ubiquitin ligase architecture, targeting, and activation. Cell 147: 10241039. Scrima A, Konickova R, Czyzewski BK, Kawasaki Y, Jeffrey PD, et al. Structural basis of UV DNA-damage recognition by the DDB1-DDB2 complicated. Cell 135: 12131223. Groisman R, Polanowska J, Kuraoka I, Sawada J, Saijo M, et al. The ubiquitin ligase activity within the DDB2 and CSA complexes is differentially regulated by the COP9 signalosome in response to DNA harm. Cell 113: 357367. Sugasawa K, Okuda Y, Saijo M, Nishi R, Matsuda N, et al. UV-induced ubiquityla.Ysis of essential components of identified DDB2 protein sub-complexes. DDB1 and Cullin4A from the ubiquitin ligase sub-complex as well as CSN5 in the COP9 signalosome subcomplex are detected together with FLAG-DDB2. Acknowledgments We thank members with the Iglehart laboratory, and Dr. Sylvie Vincent for helpful discussions. Author Contributions Conceived and created the experiments: MD JDI JBL. Performed the experiments: MD ASC JG DJE MRS GLF. Analyzed the information: MD 1676428 JBL ASC JG DS. Contributed reagents/materials/analysis tools: SI. Wrote the paper: MD JDI JBL DEF SI DS. References 1. Jackson SP, Bartek J The DNA-damage response in human biology and disease. Nature 461: 10711078. two. Keeney S, Chang GJ, Linn S Characterization of a human DNA damage binding protein implicated in xeroderma pigmentosum E. J Biol Chem 268: 2129321300. 3. Takao M, Abramic M, Moos M Jr, Otrin VR, Wootton JC, et al. A 127 kDa element of a UV-damaged DNA-binding complex, which can be defective in some xeroderma pigmentosum group E patients, is homologous to a slime mold protein. Nucleic Acids Res 21: 41114118. 4. Vermeulen W Dynamics of mammalian NER proteins. DNA Repair ten: 760771. five. Curtin NJ DNA repair dysregulation from cancer driver to therapeutic target. Nat Rev Cancer 12: 801817. 6. Ravanat JL, Douki T, Cadet J Direct and indirect effects of UV radiation on DNA and its elements. J Photochem Photobiol B 63: 88102. 7. Cadet J, Sage E, Douki T Ultraviolet radiation-mediated harm to cellular DNA. Mutat Res 571: 317. eight. Tang JY, Hwang BJ, Ford JM, Hanawalt Computer, Chu G Xeroderma pigmentosum p48 gene enhances global genomic repair and suppresses UVinduced mutagenesis. Mol Cell 5: 737744. 9. Chu G, Chang E Xeroderma pigmentosum group E cells lack a nuclear element that binds to damaged DNA. Science 242: 564567. ten. Alekseev S, Kool H, Rebel H, Fousteri M, Moser J, et al. Enhanced DDB2 expression protects mice from carcinogenic effects of chronic UV-B irradiation. Cancer Res 65: 1029810306. 11. Moser J, Volker M, Kool H, Alekseev S, Vrieling H, et al. The UVdamaged DNA binding protein mediates efficient targeting of the nucleotide 12. excision repair complex to UV-induced photo lesions. DNA Repair four: 571582. Fujiwara Y, Masutani C, Mizukoshi T, Kondo J, Hanaoka F, et al. Characterization of DNA recognition by the human UV-damaged DNAbinding protein. J Biol Chem 274: 2002720033. Reardon JT, Nichols AF, Keeney S, Smith CA, Taylor JS, et al. Comparative analysis of binding of human damaged DNA-binding protein and Escherichia coli damage recognition protein towards the key ultraviolet photoproducts: TT, TT, TT, and TT. J Biol Chem 268: 2130121308. Treiber DK, Chen Z, Essigmann JM An ultraviolet light-damaged DNA recognition protein absent in xeroderma pigmentosum group E cells binds selectively to pyrimidine pyrimidone photoproducts. Nucleic Acids Res 20: 58055810. Fischer ES, Scrima A, Bohm K, Matsumoto S, Lingaraju GM, et al. The molecular basis of CRL4DDB2/CSA ubiquitin ligase architecture, targeting, and activation. Cell 147: 10241039. Scrima A, Konickova R, Czyzewski BK, Kawasaki Y, Jeffrey PD, et al. Structural basis of UV DNA-damage recognition by the DDB1-DDB2 complex. Cell 135: 12131223. Groisman R, Polanowska J, Kuraoka I, Sawada J, Saijo M, et al. The ubiquitin ligase activity inside the DDB2 and CSA complexes is differentially regulated by the COP9 signalosome in response to DNA damage. Cell 113: 357367. Sugasawa K, Okuda Y, Saijo M, Nishi R, Matsuda N, et al. UV-induced ubiquityla.