Critical role for chicken Rad17 and Rad9 in the cellular response to DNA damage and stalled DNA replication.
The Rad17-replication factor C (Rad17-RFC) and Rad9-Rad1-Hus1 complexes are thought to function in the early phase of cell-cycle checkpoint control as sensors for genome damage and genome replication errors. However, genetic analysis of the functions of these complexes in vertebrates is complicated by the lethality of these gene disruptions in embryonic mouse cells. We disrupted the Rad17 and Rad9 loci by gene targeting in the chicken B lymphocyte line DT40. Rad17-/- and Rad9-/- DT40 cells are viable, and are highly sensitive to UV irradiation, alkylating agents, and DNA replication inhibitors, such as hydroxyurea. We further found that Rad17-/- and Rad9-/- but not ATM-/- cells are defective in S-phase DNA damage checkpoint controls and in the cellular response to stalled DNA replication. These results indicate a critical role for chicken Rad17 and Rad9 in the cellular response to stalled DNA replication and DNA damage.
Kobayashi
Masahiko
M
Department of Molecular Pathology, Cancer Research Institute, Kanazawa University, Ishikawa 920-0934, Japan.
Hirano
Atsushi
A
Kumano
Tomoyasu
T
Xiang
Shuang-Lin
SL
Mihara
Keiko
K
Haseda
Yasunari
Y
Matsui
Osamu
O
Shimizu
Hiroko
H
Yamamoto
Ken-ichi
K
D000595:Amino Acid Sequence; D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002645:Chickens; D004249:DNA Damage / Q000502:physiology*; D004261:DNA Replication / Q000502:physiology*; D004268:DNA-Binding Proteins; D006918:Hydroxyurea / Q000494:pharmacology; D008969:Molecular Sequence Data; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D016196:S Phase / Q000502:physiology; D013997:Time Factors; D014466:Ultraviolet Rays; D014965:X-Rays
D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D019384:Nucleic Acid Synthesis Inhibitors; C069930:rad9 protein; D006918:Hydroxyurea