TY - JOUR
T1 - A FancD2-monoubiquitin fusion reveals hidden functions of Fanconi anemia core complex in DNA repair
AU - Matsushita, Nobuko
AU - Kitao, Hiroyuki
AU - Ishiai, Masamichi
AU - Nagashima, Naoki
AU - Hirano, Seiki
AU - Okawa, Katsuya
AU - Ohta, Tomohiko
AU - Yu, David S.
AU - McHugh, Peter J.
AU - Hickson, Ian D.
AU - Venkitaraman, Ashok R.
AU - Kurumizaka, Hitoshi
AU - Takata, Minoru
N1 - Funding Information:
We would like to thank Dr. Larry H. Thompson (Lawrence Livermore National Laboratory) for critical reading of the manuscript, Prof. Kenshi Komatsu (Radiation Biology Centre, Kyoto University) for anti-chicken FancD2 antibody, Dr. Teru Kanda for plasmid, Mr. Taichi Shirao (Leica Microsystems) for advice in confocal microscopy, Drs. Mioko Ohzeki and Sohsuke Seki (Kawasaki Medical School) for providing DT40 mutants, Ms. Masayo Kimura and Keiko Namikoshi for expert technical help, and Ms. Kazuko Hikasa and Kyoko Takahashi for secretarial assistance. This work was supported in part by Grants-in-Aid from the Ministry of Education, Science, Sports, and Culture of Japan (M.T.). Financial supports were also provided by The Naito Foundation, The Sagawa Foundation for Promotion of Cancer Research, Kawasaki Medical School (Project Research Grant 15-201A, 16-205T, and 16-206T), and Cancer Research UK.
PY - 2005/9/16
Y1 - 2005/9/16
N2 - In DNA damage responses, the Fanconi anemia (FA) protein, FancD2, is targeted to chromatin and forms nuclear foci following its monoubiquitination, a process likely catalyzed by the FA core complex. Here, we show that a chicken FancD2-ubiquitin fusion protein, carrying a Lys-Arg substitution removing the natural monoubiquitination site (D2KR-Ub), could reverse cisplatin hypersensitivity and localize to chromatin in FANCD2-deficient DT40 cells. Importantly, the chromatin targeting was dependent on three core complex components as well as the hydrophobic surface of ubiquitin that may direct protein-protein interactions. Furthermore, a constitutively chromatin bound fusion of D2KR-histone H2B could complement cisplatin sensitivity in FANCD2- but not FANCC-, FANCG-, or FANCL-deficient cells. Thus these core complex components have an additional function in the DNA repair, which is independent of the monoubiquitination and chromatin targeting of FancD2. These results define functional consequences of FancD2 monoubiquitination and reveal previously hidden functions for the FA protein core complex.
AB - In DNA damage responses, the Fanconi anemia (FA) protein, FancD2, is targeted to chromatin and forms nuclear foci following its monoubiquitination, a process likely catalyzed by the FA core complex. Here, we show that a chicken FancD2-ubiquitin fusion protein, carrying a Lys-Arg substitution removing the natural monoubiquitination site (D2KR-Ub), could reverse cisplatin hypersensitivity and localize to chromatin in FANCD2-deficient DT40 cells. Importantly, the chromatin targeting was dependent on three core complex components as well as the hydrophobic surface of ubiquitin that may direct protein-protein interactions. Furthermore, a constitutively chromatin bound fusion of D2KR-histone H2B could complement cisplatin sensitivity in FANCD2- but not FANCC-, FANCG-, or FANCL-deficient cells. Thus these core complex components have an additional function in the DNA repair, which is independent of the monoubiquitination and chromatin targeting of FancD2. These results define functional consequences of FancD2 monoubiquitination and reveal previously hidden functions for the FA protein core complex.
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U2 - 10.1016/j.molcel.2005.08.018
DO - 10.1016/j.molcel.2005.08.018
M3 - Article
C2 - 16168378
AN - SCOPUS:24944461145
SN - 1097-2765
VL - 19
SP - 841
EP - 847
JO - Molecular Cell
JF - Molecular Cell
IS - 6
ER -