The recruitment of DNA ligase I to replication foci in S phase depends on a replication factory targeting sequence that also mediates the interaction with proliferating cell nuclear antigen (PCNA) in vitro. By exploiting a monoclonal antibody directed at a phospho-epitope, we demonstrate that Ser66 of DNA ligase I, which is part of a strong CKII consensus site, is phosphorylated in a cell cycle-dependent manner. After dephosphorylation in early G(1), the level of Ser66 phosphorylation is minimal in G(1), increases progressively in S and peaks in G(2)/M phase. The analysis of epitope-tagged DNA ligase I mutants demonstrates that dephosphorylation of Ser66 requires both the nuclear localization and the PCNA-binding site of the enzyme. Finally, we show that DNA ligase I and PCNA interact in vivo in G(1) and S phase but not in G(2)/M. We propose that dephosphorylation of Ser66 is part of a novel control mechanism to establish the pre-replicative form of DNA ligase I.

Rossi, R., Villa, A., Negri, C., Scovassi, I., Ciarrocchi, G., Biamonti, G., et al. (1999). The replication factory targeting sequence/PCNA-binding site is required in G1 to control the phosphorylation status of DNA ligase I. EMBO JOURNAL, 18(20), 5745-5754 [10.1093/emboj/18.20.5745].

The replication factory targeting sequence/PCNA-binding site is required in G1 to control the phosphorylation status of DNA ligase I

VILLA, ANTONELLO;
1999

Abstract

The recruitment of DNA ligase I to replication foci in S phase depends on a replication factory targeting sequence that also mediates the interaction with proliferating cell nuclear antigen (PCNA) in vitro. By exploiting a monoclonal antibody directed at a phospho-epitope, we demonstrate that Ser66 of DNA ligase I, which is part of a strong CKII consensus site, is phosphorylated in a cell cycle-dependent manner. After dephosphorylation in early G(1), the level of Ser66 phosphorylation is minimal in G(1), increases progressively in S and peaks in G(2)/M phase. The analysis of epitope-tagged DNA ligase I mutants demonstrates that dephosphorylation of Ser66 requires both the nuclear localization and the PCNA-binding site of the enzyme. Finally, we show that DNA ligase I and PCNA interact in vivo in G(1) and S phase but not in G(2)/M. We propose that dephosphorylation of Ser66 is part of a novel control mechanism to establish the pre-replicative form of DNA ligase I.
Articolo in rivista - Articolo scientifico
G1 Phase; 3T3 Cells; Cell Line; Hela Cells; Epitopes; Proliferating Cell Nuclear Antigen; Serine; Animals; Binding Sites; Humans; COS Cells; Mice; S Phase; DNA Replication; Phosphorylation; Antibodies, Monoclonal; DNA Ligases
English
15-ott-1999
18
20
5745
5754
none
Rossi, R., Villa, A., Negri, C., Scovassi, I., Ciarrocchi, G., Biamonti, G., et al. (1999). The replication factory targeting sequence/PCNA-binding site is required in G1 to control the phosphorylation status of DNA ligase I. EMBO JOURNAL, 18(20), 5745-5754 [10.1093/emboj/18.20.5745].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/18444
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