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Activation of Rad53 kinase in response to DNA damage and its effect in modulating phosphorylation of the lagging strand DNA polymerase
Author(s) -
Pellicioli Achille,
Lucca Chiara,
Liberi Giordano,
Marini Federica,
Lopes Massimo,
Plevani Paolo,
Romano Alfredo,
Di Fiore Pier Paolo,
Foiani Marco
Publication year - 1999
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/18.22.6561
Subject(s) - g2 m dna damage checkpoint , checkpoint kinase 2 , biology , autophosphorylation , microbiology and biotechnology , dna damage , chek1 , dna replication , dna polymerase , cell cycle checkpoint , phosphorylation , protein kinase a , cell cycle , dna , biochemistry , protein serine threonine kinases , gene
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 G 1 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.