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Yeast DNA ligase IV mediates non-homologous DNA end joining

Nature volume 388, pages 495–498 (1997)Cite this article

Abstract

The discovery of homologues from the yeast Saccharomyces cerevisiae of the human Ku DNA-end-binding proteins (HDF1 and KU80) has established that this organism is capable of non-homologous double-strand end joining (NHEJ)1,2,3,4,5, a form of DNA double-strand break repair (DSBR) active in mammalian V(D)J recombination6,7,8. Identification of the DNA ligase that mediates NHEJ in yeast will help elucidate the function of the four mammalian DNA ligases in DSBR, V(D)J recombination and other reactions9,10. Here we show that S. cerevisiae has two typical DNA ligases, the known DNA ligase I homologue CDC9 (refs 11,12, 13, 14) and the previously unknown DNA ligase IV homologue DNL4. dnl4 mutants are deficient in precise and end-processed NHEJ. DNL4 and HDF1 are epistatic in this regard, with the mutation of each having equivalent effects. dn14 mutants are complemented by overexpression of Dnl4 but not of Cdc9, and deficiency of Dnl4 alone does not impair either cell growth or the Cdc9-mediated responses to ionizing and ultraviolet radiation. Thus, S.cerevisiae has two distinct and separate ligation pathways.

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Figure 1: Multiple sequence alignment of human DNA ligase I (hDNL1), yeast CDC9, human DNA ligase IV (hDNL4) and yeast DNL4.
Figure 2: GST–Dnl4 is a pre-adenylated DNA ligase.
Figure 3: Dnl4 is required for NHEJ but not for excision or homologous DNA repair.
Figure 4: Imprecise NHEJ events in HDF1/DNL4 yeast show microdeletion and end-filling.

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Acknowledgements

We thank M. Johnston for reagents for yeast-gene disruption, and for helpful discussion on the design of these experiments. T.E.W. is a Howard Hughes Medical Institute physician postdoctoral fellow. This research is supported by NIH grants to M.R.L. M.R.L. is a Leukemia Society of America Scholar.

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  1. Ulf Grawunder & Michael R. Lieber

    Present address: Norris Comprehensive Cancer Center, University of Southern California, Rm 5425, Mailstop 73, 1441 Eastlake Avenue, Los Angeles, California, 90033, USA

Authors and Affiliations

  1. Division of Laboratory Medicine, Washington University School of Medicine, Box 8118, 660 S. Euclid Avenue, St Louis, 63110, Missouri, USA

    Thomas E. Wilson

  2. Division of Molecular Oncology, Department of Pathology, Box 8118, 660 S. Euclid Avenue, St Louis, 63110, Missouri, USA

    Thomas E. Wilson, Ulf Grawunder & Michael R. Lieber

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Correspondence to Michael R. Lieber.

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Wilson, T., Grawunder, U. & Lieber, M. Yeast DNA ligase IV mediates non-homologous DNA end joining. Nature 388, 495–498 (1997). https://doi.org/10.1038/41365

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