KIAA1018/FAN1 nuclease protects cells against genomic instability induced by interstrand cross-linking agents.
Fanconi anemia (FA) is a rare genetic disease characterized by congenital defects, bone marrow failure, chromosomal instability, and cancer susceptibility. One hallmark of cells from FA patients is hypersensitivity to interstrand cross-linking agents, such as the chemotherapeutics cisplatin and mitomycin C (MMC). We have recently characterized a FANCD2/FANCI-associated nuclease, KIAA1018/FAN1, the depletion of which sensitizes human cells to these agents. However, as the down-regulation of FAN1 in human cells was mediated by siRNA and thus only transient, we were unable to study the long-term effects of FAN1 loss on chromosomal stability. We now describe the generation of chicken DT40 B cells, in which the FAN1 locus was disrupted by gene targeting. FAN1-null cells are highly sensitive to cisplatin and MMC, but not to ionizing or UV radiation, methyl methanesulfonate, or camptothecin. The cells do not display elevated sister chromatid exchange frequencies, either sporadic or MMC-induced. Interestingly, MMC treatment causes chromosomal instability that is quantitatively, but not qualitatively, comparable to that seen in FA cells. This finding, coupled with evidence showing that DT40 cells deficient in both FAN1 and FANCC, or FAN1 and FANCJ, exhibited increased sensitivity to cisplatin compared with cells lacking only FAN1, suggests that, despite its association with FANCD2/FANCI, FAN1 in DT40 cells participates in the processing of damage induced by interstrand cross-linking-generating agents also independently of the classical FA pathway.
Department of Radiation Genetics, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
Kazunori
K
Yoshikiyo
Katja
K
Kratz
Kouji
K
Hirota
Kana
K
Nishihara
Minoru
M
Takata
Hitoshi
H
Kurumizaka
Satoshi
S
Horimoto
Shunichi
S
Takeda
Josef
J
Jiricny
D000818:Animals; D002460:Cell Line; D002645:Chickens; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004247:DNA / Q000187:drug effects*; D004249:DNA Damage; D004260:DNA Repair; D004706:Endodeoxyribonucleases; D005090:Exodeoxyribonucleases / Q000235:genetics / Q000378:metabolism*; D005199:Fanconi Anemia / Q000235:genetics; D052236:Fanconi Anemia Complementation Group D2 Protein / Q000235:genetics / Q000378:metabolism; D055785:Gene Knockdown Techniques; D042822:Genomic Instability / Q000187:drug effects*; D006801:Humans; D064251:Multifunctional Enzymes
D003432:Cross-Linking Reagents; D052236:Fanconi Anemia Complementation Group D2 Protein; D064251:Multifunctional Enzymes; D004247:DNA; D004706:Endodeoxyribonucleases; D005090:Exodeoxyribonucleases; C551063:FAN1 protein, human