Direct kinase-to-kinase signaling mediated by the FHA phosphoprotein recognition domain of the Dun1 DNA damage checkpoint kinase

Vladimir I. Bashkirov, Elena V. Bashkirova, Edwin Haghnazari, Wolf Dietrich Heyer

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

The serine-threonine kinase Dun1 contains a forkhead-associated (FHA) domain and functions in the DNA damage checkpoint pathway of Saccharomyces cerevisiae. It belongs to the Chk2 family of checkpoint kinases, which includes S. cerevisiae Rad53 and Mek1, Schizosaccharomyces pombe Cds1, and human Chk2. Dun1 is required for DNA damage-induced transcription of certain target genes, transient G2/M arrest after DNA damage, and DNA damage-induced phosphorylation of the DNA repair protein Rad55. Here we report that the FHA phosphoprotein recognition domain of Dun1 is required for direct phosphorylation of Dun1 by Rad53 kinase in vitro and in vivo. trans phosphorylation by Rad53 does not require the Dun1 kinase activity and is likely to involve only a transient interaction between the two kinases. The checkpoint functions of Dun1 kinase in DNA damage-induced transcription, G2/M cell cycle arrest, and Rad55 phosphorylation are severely compromised in an FHA domain mutant of Dun1. As a consequence, the Dun1 FHA domain mutant displays enhanced sensitivity to genotoxic stress induced by UV, methyl methanesulfonate, and the replication inhibitor hydroxyurea. We show that the Dunl FHA domain is critical for direct kinase-to-kinase signaling from Rad53 to Dun1 in the DNA damage checkpoint pathway.

Original languageEnglish (US)
Pages (from-to)1441-1452
Number of pages12
JournalMolecular and Cellular Biology
Volume23
Issue number4
DOIs
StatePublished - Feb 2003

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Phosphoproteins
DNA Damage
Phosphotransferases
Phosphorylation
Saccharomyces cerevisiae
G2 Phase Cell Cycle Checkpoints
Methyl Methanesulfonate
Hydroxyurea
Schizosaccharomyces
Protein-Serine-Threonine Kinases
DNA Repair
Genes
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Direct kinase-to-kinase signaling mediated by the FHA phosphoprotein recognition domain of the Dun1 DNA damage checkpoint kinase. / Bashkirov, Vladimir I.; Bashkirova, Elena V.; Haghnazari, Edwin; Heyer, Wolf Dietrich.

In: Molecular and Cellular Biology, Vol. 23, No. 4, 02.2003, p. 1441-1452.

Research output: Contribution to journalArticle

Bashkirov, Vladimir I. ; Bashkirova, Elena V. ; Haghnazari, Edwin ; Heyer, Wolf Dietrich. / Direct kinase-to-kinase signaling mediated by the FHA phosphoprotein recognition domain of the Dun1 DNA damage checkpoint kinase. In: Molecular and Cellular Biology. 2003 ; Vol. 23, No. 4. pp. 1441-1452.
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