Abstract
The Saccharomyces cerevisiae Rad53 protein kinase is required for the execution of checkpoint arrest at multiple stages of the cell cycle. We found that Rad53 autophosphorylation activity depends on in trans phosphorylation mediated by Mec1 and does not require physical association with other proteins. Uncoupling in trans phosphorylation from autophosphorylation using a rad53 kinase-defective mutant results in a dominant-negative checkpoint defect. Activation of Rad53 in response to DNA damage in G1 requires the Rad9, Mec3, Ddc1, Rad17 and Rad24 checkpoint factors, while this dependence is greatly reduced in S phase cells. Furthermore, during recovery from checkpoint activation, Rad53 activity decreases through a process that does not require protein synthesis. We also found that Rad53 modulates the lagging strand replication apparatus by controlling phosphorylation of the DNA polymerase α-primase complex in response to intra-S DNA damage.
| Original language | English |
|---|---|
| Pages (from-to) | 6561-6572 |
| Number of pages | 12 |
| Journal | EMBO Journal |
| Volume | 18 |
| Issue number | 22 |
| DOIs | |
| Publication status | Published - Nov 15 1999 |
Fingerprint
Keywords
- Checkpoints
- DNA damage
- DNA polymerase α-primase
- DNA replication
- Rad53
ASJC Scopus subject areas
- Genetics
- Cell Biology
Cite this
Activation of Rad53 kinase in response to DNA damage and its effect in modulating phosphorylation of the lagging strand DNA polymerase. / Pellicioli, Achille; Lucca, Chiara; Liberi, Giordano; Marini, Federica; Lopes, Massimo; Plevani, Paolo; Romano, Alfredo; Di Fiore, Pier Paolo; Foiani, Marco.
In: EMBO Journal, Vol. 18, No. 22, 15.11.1999, p. 6561-6572.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Activation of Rad53 kinase in response to DNA damage and its effect in modulating phosphorylation of the lagging strand DNA polymerase
AU - Pellicioli, Achille
AU - Lucca, Chiara
AU - Liberi, Giordano
AU - Marini, Federica
AU - Lopes, Massimo
AU - Plevani, Paolo
AU - Romano, Alfredo
AU - Di Fiore, Pier Paolo
AU - Foiani, Marco
PY - 1999/11/15
Y1 - 1999/11/15
N2 - The Saccharomyces cerevisiae Rad53 protein kinase is required for the execution of checkpoint arrest at multiple stages of the cell cycle. We found that Rad53 autophosphorylation activity depends on in trans phosphorylation mediated by Mec1 and does not require physical association with other proteins. Uncoupling in trans phosphorylation from autophosphorylation using a rad53 kinase-defective mutant results in a dominant-negative checkpoint defect. Activation of Rad53 in response to DNA damage in G1 requires the Rad9, Mec3, Ddc1, Rad17 and Rad24 checkpoint factors, while this dependence is greatly reduced in S phase cells. Furthermore, during recovery from checkpoint activation, Rad53 activity decreases through a process that does not require protein synthesis. We also found that Rad53 modulates the lagging strand replication apparatus by controlling phosphorylation of the DNA polymerase α-primase complex in response to intra-S DNA damage.
AB - The Saccharomyces cerevisiae Rad53 protein kinase is required for the execution of checkpoint arrest at multiple stages of the cell cycle. We found that Rad53 autophosphorylation activity depends on in trans phosphorylation mediated by Mec1 and does not require physical association with other proteins. Uncoupling in trans phosphorylation from autophosphorylation using a rad53 kinase-defective mutant results in a dominant-negative checkpoint defect. Activation of Rad53 in response to DNA damage in G1 requires the Rad9, Mec3, Ddc1, Rad17 and Rad24 checkpoint factors, while this dependence is greatly reduced in S phase cells. Furthermore, during recovery from checkpoint activation, Rad53 activity decreases through a process that does not require protein synthesis. We also found that Rad53 modulates the lagging strand replication apparatus by controlling phosphorylation of the DNA polymerase α-primase complex in response to intra-S DNA damage.
KW - Checkpoints
KW - DNA damage
KW - DNA polymerase α-primase
KW - DNA replication
KW - Rad53
UR - http://www.scopus.com/inward/record.url?scp=0033570894&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033570894&partnerID=8YFLogxK
U2 - 10.1093/emboj/18.22.6561
DO - 10.1093/emboj/18.22.6561
M3 - Article
C2 - 10562568
AN - SCOPUS:0033570894
VL - 18
SP - 6561
EP - 6572
JO - EMBO Journal
JF - EMBO Journal
SN - 0261-4189
IS - 22
ER -