Activation of Rad53 kinase in response to DNA damage and its effect in modulating phosphorylation of the lagging strand DNA polymerase

Achille Pellicioli, Chiara Lucca, Giordano Liberi, Federica Marini, Massimo Lopes, Paolo Plevani, Alfredo Romano, Pier Paolo Di Fiore, Marco Foiani

Research output: Contribution to journalArticle

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 languageEnglish
Pages (from-to)6561-6572
Number of pages12
JournalEMBO Journal
Volume18
Issue number22
DOIs
Publication statusPublished - Nov 15 1999

Fingerprint

Phosphorylation
DNA-Directed DNA Polymerase
DNA Damage
Phosphotransferases
Chemical activation
DNA
DNA Primase
Saccharomyces cerevisiae Proteins
S Phase
Yeast
Protein Kinases
Cell Cycle
Proteins
Cells
Association reactions
Recovery
Defects

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 journalArticle

Pellicioli, A, Lucca, C, Liberi, G, Marini, F, Lopes, M, Plevani, P, Romano, A, Di Fiore, PP & Foiani, M 1999, 'Activation of Rad53 kinase in response to DNA damage and its effect in modulating phosphorylation of the lagging strand DNA polymerase', EMBO Journal, vol. 18, no. 22, pp. 6561-6572. https://doi.org/10.1093/emboj/18.22.6561
Pellicioli, Achille ; Lucca, Chiara ; Liberi, Giordano ; Marini, Federica ; Lopes, Massimo ; Plevani, Paolo ; Romano, Alfredo ; Di Fiore, Pier Paolo ; Foiani, Marco. / Activation of Rad53 kinase in response to DNA damage and its effect in modulating phosphorylation of the lagging strand DNA polymerase. In: EMBO Journal. 1999 ; Vol. 18, No. 22. pp. 6561-6572.
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