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Journal articles
Surprising complexity of the Asf1 histone chaperone-Rad53 kinase interaction
Abstract : The histone chaperone Asf1 and the checkpoint kinase Rad53 are found in a complex in budding yeast cells in the absence of genotoxic stress. Our data suggest that this complex involves at least three interaction sites. One site involves the H3-binding surface of Asf11 with an as yet undefined surface of Rad53. A second site is formed by the Rad53-FHA1 domain binding to Asf1-$T_{270}$ phosphorylated by casein kinase II. The third site involves the C-terminal 21 amino acids of Rad53 bound to the conserved Asf1 N-terminal domain. The structure of this site showed that the Rad53 C-terminus binds Asf1 in a remarkably similar manner to peptides derived from the histone cochaperones HirA and CAF-I. We call this binding motif, $(R/K)R(I/A/V)$x$(L/P)$, the AIP box for Asf1-Interacting Protein box. Furthermore, C-terminal Rad53-$F_{820}$ binds the same pocket of Asf1 as does histone $H4-F_{100}$. Thus Rad53 competes with histones H3-H4 and cochaperones HirA/CAF-I for binding to Asf1. Rad53 is phosphorylated and activated upon genotoxic stress. The Asf1-Rad53 complex dissociated when cells were treated with hydroxyurea but not methyl-methane-sulfonate, suggesting a regulation of the complex as a function of the stress. We identified a rad53 mutation that destabilized the Asf1-Rad53 complex and increased the viability of rad9 and rad24 mutants in conditions of genotoxic stress, suggesting that complex stability impacts the DNA damage response.
https://hal-cea.archives-ouvertes.fr/cea-01744552
Contributor : Florence Mousson <>
Submitted on : Tuesday, March 27, 2018 - 2:44:31 PM
Last modification on : Monday, December 14, 2020 - 4:52:21 PM
Contributor : Florence Mousson <>
Submitted on : Tuesday, March 27, 2018 - 2:44:31 PM
Last modification on : Monday, December 14, 2020 - 4:52:21 PM
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