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Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
Interplay of DNA damage and cell cycle signaling at the level of human replication protein A.
DNA Repair 21, 12-23 (2014)
Replication protein A (RPA) is the main human single-stranded DNA (ssDNA)-binding protein. It is essential for cellular DNA metabolism and has important functions in human cell cycle and DNA damage signaling. RPA is indispensable for accurate homologous recombination (HR)-based DNA double-strand break (DSB) repair and its activity is regulated by phosphorylation and other post-translational modifications. HR occurs only during S and G2 phases of the cell cycle. All three subunits of RPA contain phosphorylation sites but the exact set of HR-relevant phosphorylation sites on RPA is unknown. In this study, a high resolution capillary isoelectric focusing immunoassay, used under native conditions, revealed the isoforms of the RPA heterotrimer in control and damaged cell lysates in G2. Moreover, the phosphorylation sites of chromatin-bound and cytosolic RPA in S and G2 phases were identified by western and IEF analysis with all available phosphospecific antibodies for RPA2. Strikingly, most of the RPA heterotrimers in control G2 cells are phosphorylated with 5 isoforms containing up to 7 phosphates. These isoforms include RPA2 pSer23 and pSer33. DNA damaged cells in G2 had 9 isoforms with up to 14 phosphates. DNA damage isoforms contained pSer4/8, pSer12, pThr21, pSer23, and pSer33 on RPA2 and up to 8 unidentified phosphorylation sites.
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Publikationstyp
Artikel: Journalartikel
Dokumenttyp
Wissenschaftlicher Artikel
Schlagwörter
Dna Damage Response ; Dna Double-stranded Breaks ; Dna Repair ; G2 Phase ; Homologous Recombination ; Isoelectric Focusing ; Phosphorylation; Strand Break Repair; Homologous Recombination; Ataxia-telangiectasia; Induced Hyperphosphorylation; Induced Phosphorylation; Binding-protein; S-phase; Saccharomyces-cerevisiae; Rpa Phosphorylation; Distinct Roles
ISSN (print) / ISBN
1568-7864
e-ISSN
1568-7856
Zeitschrift
DNA Repair
Quellenangaben
Band: 21,
Seiten: 12-23
Verlag
Elsevier
Verlagsort
Amsterdam
Nichtpatentliteratur
Publikationen
Begutachtungsstatus
Peer reviewed
Institut(e)
Institute of Molecular Immunology (IMI)