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Impact of DNA ligase IV on the fidelity of end joining in human cells. | LitMetric

AI Article Synopsis

  • A study on LIG4-null human pre-B cell lines reveals that these cells struggle with effective DNA end joining, showcasing the essential role of DNA ligase IV in this process.
  • Cells with hypomorphic mutations in LIG4 maintain some activity but still display reduced fidelity during DNA rejoining, indicating that even partial function can lead to errors.
  • Additionally, DNA ligase IV, in combination with XRCC4, helps protect DNA ends from degradation, which may enhance the accuracy of DNA end rejoining in living cells.

Article Abstract

A DNA ligase IV (LIG4)-null human pre-B cell line and human cell lines with hypomorphic mutations in LIG4 are significantly impaired in the frequency and fidelity of end joining using an in vivo plasmid assay. Analysis of the null line demonstrates the existence of an error-prone DNA ligase IV-independent rejoining mechanism in mammalian cells. Analysis of lines with hypomorphic mutations demonstrates that residual DNA ligase IV activity, which is sufficient to promote efficient end joining, nevertheless can result in decreased fidelity of rejoining. Thus, DNA ligase IV is an important factor influencing the fidelity of end joining in vivo. The LIG4-defective cell lines also showed impaired end joining in an in vitro assay using cell-free extracts. Elevated degradation of the terminal nucleotide was observed in a LIG4-defective line, and addition of the DNA ligase IV-XRCC4 complex restored end protection. End protection by DNA ligase IV was not dependent upon ligation. Finally, using purified proteins, we demonstrate that DNA ligase IV-XRCC4 is able to protect DNA ends from degradation by T7 exonuclease. Thus, the ability of DNA ligase IV-XRCC4 to protect DNA ends may contribute to the ability of DNA ligase IV to promote accurate rejoining in vivo.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC153745PMC
http://dx.doi.org/10.1093/nar/gkg317DOI Listing

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