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Regulated degradation of replication-dependent histone mRNAs requires both ATR and Upf1

Nature Structural & Molecular Biology volume 12, pages 794–800 (2005)Cite this article

Abstract

Eukaryotic cells coordinately regulate histone and DNA synthesis. In mammalian cells, most of the regulation of histone synthesis occurs post-transcriptionally by regulating the concentrations of histone mRNA. As cells enter S phase, histone mRNA levels increase, and at the end of S phase they are rapidly degraded. Moreover, inhibition of DNA synthesis causes rapid degradation of histone mRNAs. Replication-dependent histone mRNAs are the only metazoan mRNAs that are not polyadenylated. Instead, they end with a conserved stem-loop structure, which is the only cis-acting element required for coupling regulation of histone mRNA half-life with DNA synthesis. Here we show that regulated degradation of histone mRNAs requires Upf1, a key regulator of the nonsense-mediated decay pathway, and ATR, a key regulator of the DNA damage checkpoint pathway activated during replication stress.

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Figure 1: siRNA-mediated downregulation of Upf1 but not Upf2 resulted in less efficient histone mRNA degradation.
Figure 2: Stable expression of dominant negative Upf1 proteins prevents efficient histone mRNA degradation after inhibition of DNA synthesis.
Figure 3: Transient overexpression of dominant negative Upf1 mutants prevents efficient histone mRNA degradation.
Figure 4: Expression of K498A mutant of Upf1 results in a delay of histone mRNA degradation as cells exited the S phase.
Figure 5: Upf1 interacts with the histone mRNP.
Figure 6: Treatment of cells with caffeine (CA) before hydroxyurea (HU) results in a decreased rate of histone mRNA degradation.
Figure 7: The kinase activity of ATR is required for efficient degradation of histone mRNAs.

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Acknowledgements

This work was supported by US National Institutes of Health grant GM29832 to W.F.M. We thank E. Wagner and B. Duronio for critical reading of the manuscript. We thank L. Maquat (University of Rochester) for Upf1 clone, M. Hentze (EMBL) and C. Smith (Cambridge University) for Upf1 antibodies, J. Lykke-Andersen (University of Colorado) for Upf2 antibody, M. Garcia-Blanco (Duke University) for PTB antibody and P. Nghiem (Harvard) for ATR cells.

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Authors and Affiliations

  1. Department of Biology, University of North Carolina, Chapel Hill, 27599, North Carolina, USA

    Handan Kaygun & William F Marzluff

  2. Program in Molecular Biology and Biotechnology, University of North Carolina, Chapel Hill, 27599, North Carolina, USA

    Handan Kaygun & William F Marzluff

  3. Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, 27599, North Carolina, USA

    William F Marzluff

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Correspondence to William F Marzluff.

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Kaygun, H., Marzluff, W. Regulated degradation of replication-dependent histone mRNAs requires both ATR and Upf1. Nat Struct Mol Biol 12, 794–800 (2005). https://doi.org/10.1038/nsmb972

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