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Replication stress checkpoint signaling controls tRNA gene transcription

Nature Structural & Molecular Biology volume 17, pages 976–981 (2010)Cite this article

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Abstract

In budding yeast, the transcriptional machinery at tRNA genes naturally interferes with replication in a way that can promote chromosome breakage. Here we show that a signaling module composed of core components of the replication stress checkpoint pathway represses this fork-pausing machinery in normally cycling and genotoxin-treated cells. Specifically, the sensor kinase Mec1, the signaling adaptor Mrc1 and the transducer kinase Rad53 relay signals that globally repress tRNA gene transcription during unchallenged proliferation and under conditions of replication stress. Repressive signaling in genotoxin-treated cells requires Rad53-dependent activation of a conserved repressor of tRNA gene transcription, Maf1. Cells lacking Maf1 are sensitive to replication stress under conditions of elevated tRNA gene transcription. We propose that checkpoint control of the fork-pausing activity of tRNA genes complements the repertoire of replisome-targeted mechanisms by which checkpoint proteins promote faithful DNA replication.

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Figure 1: Control of tRNA gene transcription by Mec1 and Mrc1 during unchallenged proliferation.
Figure 2: Control of tRNA gene transcription by transducer kinase Rad53 during unchallenged proliferation.
Figure 3: Repression of tRNA gene transcription under conditions of replication stress.
Figure 4: Maf1-dependent repression of tRNA gene transcription under conditions of replication stress.

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Acknowledgements

We are grateful to Y. Sanchez, D. Stern and I. Willis for genetic reagents. K. Robinson, D. Stuart, M. Weinfeld, M. Grenon, N. Lowndes, I. Willis, R. Moir, O. Lefebvre and L. Minard are thanked for technical advice and M. MacDonald for technical assistance. R. Wellinger, T. Weinert, P. Pasero and J. Cobb are acknowledged for helpful discussions. This work was supported by grants to M.C.S. from the National Cancer Institute of Canada (supported by the Canadian Cancer Society), the Canadian Institutes for Health Research, the Cancer Research Society and the Alberta Heritage Foundation for Medical Research (AHFMR). V.C.N. was supported by studentships from the AHFMR and the National Science and Engineering Research Council of Canada. H.E.M. was supported by a summer studentship from the AHFMR.

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  1. Department of Biochemistry, School of Molecular and Systems Medicine, University of Alberta, Edmonton, Alberta, Canada

    Vesna C Nguyen, Brett W Clelland, Darren J Hockman, Sonya L Kujat-Choy, Holly E Mewhort & Michael C Schultz

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  1. Vesna C Nguyen
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Contributions

V.C.N. generated about half of the yeast strains and performed most of the in vivo transcription experiments as well as the assays of Maf1 phosphorylation; B.W.C. generated about half the yeast strains, performed the ChIP experiments and in vivo MAF1 epistasis transcription experiments and developed the maf1Δ HU sensitivity assay; S.L.K.-C. discovered HU repression by Maf1; D.J.H. and H.E.M. provided technical assistance to V.C.N. and B.W.C.; M.C.S. conceived the project, performed the in vitro transcription experiments and wrote the manuscript.

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Correspondence to Michael C Schultz.

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Nguyen, V., Clelland, B., Hockman, D. et al. Replication stress checkpoint signaling controls tRNA gene transcription. Nat Struct Mol Biol 17, 976–981 (2010). https://doi.org/10.1038/nsmb.1857

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