DNA ligase I and Nbs1 proteins associate in a complex and colocalize at replication factories

Riccardo Vago, Valentina Leva, Giuseppe Biamonti, Alessandra Montecucco

Research output: Contribution to journalArticle

Abstract

DNA ligase I is the main DNA ligase activity involved in eukaryotic DNA replication acting in the joining of Okazaki fragments. This enzyme is also implicated in nucleotide excision repair and in the long-patch base excision repair while its role in the recombinational repair pathways is poorly understood. DNA ligase I is phosphorylated during cell cycle at several serine and threonine residues that regulate its participation in different DNA transactions by modulating the interaction with different protein partners. Here we use an antibody-based array method to identify novel DNA ligase-interacting partners. We show that DNA ligase I participates in several multiprotein complexes with proteins involved in DNA replication and repair, cell cycle control, and protein modification. In particular we demonstrate that DNA ligase I complexes with Nbs1, a core component of the MRN complex critical for detection, processing and repair of double-stranded DNA breaks. The analysis of epitope tagged DNA ligase I mutants demonstrates that the association is mediated by the catalytic fragment of the enzyme. DNA ligase I and Nbs1 colocalize at replication factories during unperturbed replication and after treatment with DNA damaging agents. Since MRN complex is involved in the repair of double-stranded DNA breaks by homologous recombination at stalled replication forks our data support the notion that DNA ligase I participates in homology dependent pathways that deal with replicationassociated lesions generated when replication fork encounters DNA damage.

Original languageEnglish
Pages (from-to)2600-2607
Number of pages8
JournalCell Cycle
Volume8
Issue number16
Publication statusPublished - Aug 15 2009

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Keywords

  • Antibody array
  • DNA ligase
  • DNA ligase I partners
  • Nbs1
  • Replication factories

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Molecular Biology

Cite this

Vago, R., Leva, V., Biamonti, G., & Montecucco, A. (2009). DNA ligase I and Nbs1 proteins associate in a complex and colocalize at replication factories. Cell Cycle, 8(16), 2600-2607.