PP4 phosphatase cooperates in recombinational DNA repair by enhancing double-strand break end resection

The role of Rad53 in response to a DNA lesion is central for the accurate orchestration of the DNA damage response. Rad53 activation relies on its phosphorylation by Mec1 and its own autophosphorylation in a manner dependent on the adaptor Rad9. While the mechanism behind Rad53 activation has been well documented, less is known about the processes that counteract its activity along the repair of a DNA adduct. Here, we describe that PP4 phosphatase is required to avoid Rad53 hyper-phosphorylation during the repair of a double-strand break, a process that impacts on the phosphorylation status of multiple factors involved in the DNA damage response. PP4-dependent Rad53 dephosphorylation stimulates DNA end resection by relieving the negative effect that Rad9 exerts over the Sgs1/Dna2 exonuclease complex. Consequently, elimination of PP4 activity affects resection and repair by single-strand annealing, defects that are bypassed by reducing Rad53 hyperphosphorylation. These results confirm that Rad53 phosphorylation is controlled by PP4 during the repair of a DNA lesion and demonstrate that the attenuation of its kinase activity during the initial steps of the repair process is essential to efficiently enhance recombinational DNA repair pathways that depend on long-range resection for their success.
Ministerio de Economía y Competitividad [BFU2013-41216-P, BFU2016-77081-P and PGC2018-097963-B-100 (MCIU/AEI/FEDER, UE) granted to A.C.-B.]; The IBFG is supported in part by an institutional grant from the ‘Junta y Castilla y León’ (Programa ‘Escalera de Excelencia’ de la Junta de Castilla y León, Ref. CLU-2017-03 cofinanciado por el P.O. FEDER de Castilla y León 14–20); M.T.V. was recipient of a predoctoral fellowship from the ‘Junta de Castilla y León’; F.R. was recipient of a predoctoral fellowship from the ‘Ministerio de Economía y Competitividad’. Funding for open access charge: Ministerio de Economía y Competitividad Grand [Reference 1: BFU2016-77081-P, Grand Reference 2: PGC2018-097963-B-100].