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NEJ1 controls non-homologous end joining in Saccharomyces cerevisiae

Nature volume 414, pages 666–669 (2001)Cite this article

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Abstract

Broken DNA ends are rejoined by non-homologous end-joining (NHEJ) pathways requiring the Ku proteins (Ku70, Ku80), DNA ligase IV and its associated protein Lif1/Xrcc4 (ref. 1). In mammalian meiotic cells, Ku protein levels are much lower than in somatic cells, apparently reducing the capacity of meiotic cells to carry out NHEJ and thereby promoting homologous recombination2. In Saccharomyces cerevisiae, NHEJ is also downregulated in meiosis-competent MATa/MATα diploid cells in comparison with diploids or haploids expressing only MATa or MATα3,4. Diploids expressing both MATa and MATα show enhanced mitotic homologous recombination4. Here we report that mating-type-dependent regulation of NHEJ in budding yeast is caused in part by transcriptional repression of both LIF1 and the gene NEJ1 (YLR265C)—identified from microarray screening of messenger RNAs. Deleting NEJ1 reduces NHEJ 100-fold in MATa or MATα haploids. Constitutive expression of NEJ1, but not expression of LIF1, restores NHEJ in MATa/MATα cells. Nej1 regulates the subcellular distribution of Lif1. A green fluorescent protein (GFP)–Lif1 fusion protein accumulates in the nucleus in cells expressing NEJ1 but is largely cytoplasmic when NEJ1 is repressed.

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Figure 1: Cell-type regulation of LIF1 and NEJ1 mRNA.
Figure 2: Effect of LIF1 and NEJ1 on NHEJ ligation of linearized plasmid DNA transformed into yeast.
Figure 3: Role of NEJ1 in chromosomal and plasmid NHEJ.
Figure 4: Localization of EGFP–Lif1 in haploid MATα and diploid MAT a/MATα cells.

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Acknowledgements

We are grateful for collaboration with A. Gasch, P. Brown and D. Botstein to analyse mRNA abundance under various conditions, and for their generosity in declining to be authors on this paper. S.E.L. is a Hildegarde A. Becher Foundation fellow of The Leukemia and Lymphoma Society. G.H. was supported by the Deutsche Forschungsgesellschaft (DFG). This work was supported by a grant of the Swiss National Science Foundation to M.B. and P.S. and DOE and National Institutes of Health grants to J.E.H.

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Author notes

  1. Gernot Herrmann

    Present address: Department of Dermatology, University of Cologne, D-50931, Cologne, Germany

  2. Sang Eun Lee

    Present address: Department of Molecular Medicine/IBT UTHSC-SA, San Antonio, Texas, 78245, USA

  3. Maria Valencia, Marc Bentele, Gernot Herrmann and Primo Schär: These authors contributed equally to this work

Authors and Affiliations

  1. Rosenstiel Center and Department of Biology, Brandeis University, Waltham, 02454-9110, Massachusetts, USA

    Maria Valencia, Moreshwar B. Vaze, Eliayhu Kraus, Sang Eun Lee & James E. Haber

  2. Institute of Medical Radiobiology, University of Zürich, Zürich, CH-8008, Switzerland

    Maria Valencia & Marc Bentele

  3. Department of Dermatology, University of Cologne, D-50931, Cologne, Germany

    Sang Eun Lee

  4. Department of Molecular Medicine/IBT UTHSC-SA, San Antonio, Texas, 78245, USA

    Gernot Herrmann

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Correspondence to James E. Haber.

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Valencia, M., Bentele, M., Vaze, M. et al. NEJ1 controls non-homologous end joining in Saccharomyces cerevisiae. Nature 414, 666–669 (2001). https://doi.org/10.1038/414666a

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