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Molecular and Cellular Biology, August 2000, p. 5888-5896, Vol. 20, No. 16
Division of Biological Science, Graduate
School of Science, Nagoya University, Chikusa-ku, Nagoya 464-0814, Japan
Received 7 February 2000/Returned for modification 20 March
2000/Accepted 2 May 2000
RAD24 and RFC5 are required for DNA damage
checkpoint control in the budding yeast Saccharomyces
cerevisiae. Rad24 is structurally related to replication factor C
(RFC) subunits and associates with RFC subunits Rfc2, Rfc3,
Rfc4, and Rfc5. rad24
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Rfc5, in Cooperation with Rad24, Controls DNA
Damage Checkpoints throughout the Cell Cycle in
Saccharomyces cerevisiae

mutants are defective in all the
G1-, S-, and G2/M-phase DNA damage checkpoints, whereas the rfc5-1 mutant is impaired only in the S-phase
DNA damage checkpoint. Both the RFC subunits and Rad24 contain a
consensus sequence for nucleoside triphosphate (NTP) binding. To
determine whether the NTP-binding motif is important for Rad24
function, we mutated the conserved lysine115 residue in
this motif. The rad24-K115E mutation, which changes lysine
to glutamate, confers a complete loss-of-function phenotype, while the rad24-K115R mutation, which changes lysine to
arginine, shows no apparent phenotype. Although neither
rfc5-1 nor rad24-K115R single mutants are
defective in the G1- and G2/M-phase DNA damage checkpoints, rfc5-1 rad24-K115R double mutants become
defective in these checkpoints. Coimmunoprecipitation experiments
revealed that Rad24K115R fails to interact with the RFC
proteins in rfc5-1 mutants. Together, these results
indicate that RFC5, like RAD24, functions in
all the G1-, S- and G2/M-phase DNA damage
checkpoints and suggest that the interaction of Rad24 with the RFC
proteins is essential for DNA damage checkpoint control.
*
Corresponding author. Mailing address: Division of
Biological Science, Graduate School of Science, Nagoya University,
Chikusa-ku, Nagoya 464-8602, Japan. Phone: 81-52-789-2593. Fax:
81-52-789-2589. E-mail:
g44177a{at}nucc.cc.nagoya-u.ac.jp.
Present address: Tsukuba Research Institute, Banyu Pharmaceutical
Co., Ltd., Tsukuba 300-2611, Japan.
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