RNF169 and RNF168 novel substrates of DYRK1A : connecting DYRK1A to DNA-damage repair

Gene dosage alterations of the kinase DYRK1A are linked to disease in humans. To better understand DYRK1A activities an interactome analysis was performed. RNF169, an E3-ubiquitin ligase key component of the cellular response to double-strand breaks (DSBs), was found as a top nuclear interactor. The...

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Detalhes bibliográficos
Autor: Roewenstrunk, Julia Maria
Formato: tesis doctoral
Estado:Versión publicada
Fecha de publicación:2016
País:España
Recursos:CBUC, CESCA
Repositorio:TDR. Tesis Doctorales en Red
OAI Identifier:oai:www.tdx.cat:10803/565442
Acesso em linha:http://hdl.handle.net/10803/565442
Access Level:acceso abierto
Palavra-chave:DYRK1A
RNF169
RNF168
DNA-damage
Phosphorylation
Daño al DNA
Fosforilación
577
Descrição
Resumo:Gene dosage alterations of the kinase DYRK1A are linked to disease in humans. To better understand DYRK1A activities an interactome analysis was performed. RNF169, an E3-ubiquitin ligase key component of the cellular response to double-strand breaks (DSBs), was found as a top nuclear interactor. The functional characterization of this interaction has uncovered that a dedicated motif in the non-catalytic N-terminus of DYRK1A is responsible of the direct interaction with RNF169 and that this interaction is essential for the recruitment of DYRK1A to DSBs. Using a combination of mass spectrometry analysis, mutagenesis, and in vitro kinase assays several DYRK1A-dependent phosphosites have been identified in RNF169 and its paralog RNF168. Reporter-cell assays and IRIF analysis showed that DYRK1A silencing perturbs the DSB-repair pathways. In agreement, DYRK1A knockdown leads to increased radiation sensitivity. All together, the data suggest a role for DYRK1A in DSB-repair that might involve the phosphorylation of RNF168 and RNF169.