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Molecular and Cellular Biology, June 2001, p. 3725-3737, Vol. 21, No. 11
Department of Molecular Biology and
Biochemistry, Center for Advanced Biotechnology and Medicine,
Rutgers University, Piscataway, New Jersey 08854
Received 28 December 2000/Returned for modification 1 February
2001/Accepted 28 March 2001
The large subunit of replication protein A (Rpa1) consists of three
single-stranded DNA binding domains and an N-terminal domain (Rpa1N) of
unknown function. To determine the essential role of this domain we
searched for mutations that require wild-type Rpa1N for viability in
yeast. A mutation in RFC4, encoding a small subunit of
replication factor C (RFC), was found to display allele-specific interactions with mutations in the gene encoding Rpa1
(RFA1). Mutations that map to Rpa1N and confer sensitivity
to the DNA synthesis inhibitor hydroxyurea, such as
rfa1-t11, are lethal in combination with
rfc4-2. The rfc4-2 mutant itself is sensitive to hydroxyurea, and like rfc2 and rfc5 strains,
it exhibits defects in the DNA replication block and intra-S
checkpoints. RFC4 and the DNA damage checkpoint gene
RAD24 were found to be epistatic with respect to DNA damage
sensitivity. We show that the rfc4-2 mutant is defective in
the G1/S DNA damage checkpoint response and that both the
rfc4-2 and rfa1-t11 strains are defective in the G2/M DNA damage checkpoint. Thus, in addition to its
essential role as part of the clamp loader in DNA replication, Rfc4
plays a role as a sensor in multiple DNA checkpoint pathways. Our
results suggest that a physical interaction between Rfc4 and Rpa1N is required for both roles.
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.11.3725-3737.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Rfc4 Interacts with Rpa1 and Is Required for Both
DNA Replication and DNA Damage Checkpoints in Saccharomyces
cerevisiae
*
Corresponding author. Mailing address: Department of
Molecular Biology and Biochemistry, Rutgers University, 679 Hoes La., CABM, Piscataway, NJ 08854. Phone: (732) 235-4197. Fax: (732) 235-4880. E-mail: brill{at}mbcl.rutgers.edu.
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