In vitro and in vivo interactions of DNA ligase IV with a subunit of the condensin complex

Marcin R. Przewloka, Paige E. Pardington, Steven M. Yannone, David J. Chen, Robert B. Cary

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Several findings have revealed a likely role for DNA ligase IV, and interacting protein XRCC4, in the final steps of mammalian DNA double-strand break repair. Recent evidence suggests that the human DNA ligase IV protein plays a critical role in the maintenance of genomic stability. To identify protein-protein interactions that may shed further light on the molecular mechanisms of DSB repair and the biological roles of human DNA ligase IV, we have used the yeast two-hybrid system in conjunction with traditional biochemical methods. These efforts have resulted in the identification of a physical association between the DNA ligase IV polypeptide and the human condensin subunit known as hCAP-E. The hCAP-E polypeptide, a member of the Structural Maintenance of Chromosomes (SMC) super-family of proteins, coimmunoprecipitates from cell extracts with DNA ligase IV. Immunofluorescence studies reveal colocalization of DNA ligase IV and hCAP-E in the interphase nucleus, whereas mitotic cells display colocalization of both polypeptides on mitotic chromosomes. Strikingly, the XRCC4 protein is excluded from the area of mitotic chromosomes, suggesting the formation of specialized DNA ligase IV complexes subject to cell cycle regulation. We discuss our findings in light of known and hypothesized roles for ligase IV and the condensin complex.

Original languageEnglish (US)
Pages (from-to)685-697
Number of pages13
JournalMolecular Biology of the Cell
Volume14
Issue number2
DOIs
StatePublished - Feb 1 2003

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Proteins
Chromosomes
Peptides
Maintenance
Two-Hybrid System Techniques
Double-Stranded DNA Breaks
Genomic Instability
Interphase
Ligases
Cell Extracts
Fluorescent Antibody Technique
DNA Ligase ATP
condensin complexes
In Vitro Techniques
Cell Cycle
human LIG4 protein
1,2-di-(4-sulfamidophenyl)-4-butylpyrazolidine-3,5-dione

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

In vitro and in vivo interactions of DNA ligase IV with a subunit of the condensin complex. / Przewloka, Marcin R.; Pardington, Paige E.; Yannone, Steven M.; Chen, David J.; Cary, Robert B.

In: Molecular Biology of the Cell, Vol. 14, No. 2, 01.02.2003, p. 685-697.

Research output: Contribution to journalArticle

Przewloka, Marcin R. ; Pardington, Paige E. ; Yannone, Steven M. ; Chen, David J. ; Cary, Robert B. / In vitro and in vivo interactions of DNA ligase IV with a subunit of the condensin complex. In: Molecular Biology of the Cell. 2003 ; Vol. 14, No. 2. pp. 685-697.
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