TY - JOUR
T1 - An ATR- and Cdc7-dependent DNA damage checkpoint that inhibits initiation of DNA replication
AU - Costanzo, Vincenzo
AU - Shechter, David
AU - Lupardus, Patrick J.
AU - Cimprich, Karlene A.
AU - Gottesman, Max
AU - Gautier, Jean
N1 - Funding Information:
We would like to thank Dr. J. Walter for help and advice setting up the NPE system and for Cdc7 antiserum. We would like to thank Dr. S. Waga for Dbf4 antiserum, Dr. P. Jackson for RPA antiserum, Dr. E. Lees for Dbf4 cDNA, Dr. M. Madine for MCM2 cDNA, and Dr. M. Michael for the geminin plasmid. This work is supported by ACS grant RSG CCG-103367, NIH grants (GM56781 and CA92245) to J.G, and NIH grant (GM062193) to K.A.C.
PY - 2003/1/1
Y1 - 2003/1/1
N2 - We have analyzed how single-strand DNA gaps affect DNA replication in Xenopus egg extracts. DNA lesions generated by etoposide, a DNA topoisomerase II inhibitor, or by exonuclease treatment activate a DNA damage checkpoint that blocks initiation of plasmid and chromosomal DNA replication. The checkpoint is abrogated by caffeine and requires ATR, but not ATM, protein kinase. The block to DNA synthesis is due to inhibition of Cdc7/Dbf4 protein kinase activity and the subsequent failure of Cdc45 to bind to chromatin. The checkpoint does not require pre-RC assembly but requires loading of the single-strand binding protein, RPA, on chromatin. This is the biochemical demonstration of a DNA damage checkpoint that targets Cdc7/Dbf4 protein kinase.
AB - We have analyzed how single-strand DNA gaps affect DNA replication in Xenopus egg extracts. DNA lesions generated by etoposide, a DNA topoisomerase II inhibitor, or by exonuclease treatment activate a DNA damage checkpoint that blocks initiation of plasmid and chromosomal DNA replication. The checkpoint is abrogated by caffeine and requires ATR, but not ATM, protein kinase. The block to DNA synthesis is due to inhibition of Cdc7/Dbf4 protein kinase activity and the subsequent failure of Cdc45 to bind to chromatin. The checkpoint does not require pre-RC assembly but requires loading of the single-strand binding protein, RPA, on chromatin. This is the biochemical demonstration of a DNA damage checkpoint that targets Cdc7/Dbf4 protein kinase.
UR - http://www.scopus.com/inward/record.url?scp=0037245862&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037245862&partnerID=8YFLogxK
U2 - 10.1016/S1097-2765(02)00799-2
DO - 10.1016/S1097-2765(02)00799-2
M3 - Article
C2 - 12535533
AN - SCOPUS:0037245862
SN - 1097-2765
VL - 11
SP - 203
EP - 213
JO - Molecular Cell
JF - Molecular Cell
IS - 1
ER -