Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Homeostatic response to hypoxia is regulated by the N-end rule pathway in plants

Nature volume 479, pages 415–418 (2011)Cite this article

Subjects

Abstract

Plants and animals are obligate aerobes, requiring oxygen for mitochondrial respiration and energy production. In plants, an unanticipated decline in oxygen availability (hypoxia), as caused by roots becoming waterlogged or foliage submergence, triggers changes in gene transcription and messenger RNA translation that promote anaerobic metabolism and thus sustain substrate-level ATP production1. In contrast to animals2, oxygen sensing has not been ascribed to a mechanism of gene regulation in response to oxygen deprivation in plants. Here we show that the N-end rule pathway of targeted proteolysis acts as a homeostatic sensor of severe low oxygen levels in Arabidopsis, through its regulation of key hypoxia-response transcription factors. We found that plants lacking components of the N-end rule pathway constitutively express core hypoxia-response genes and are more tolerant of hypoxic stress. We identify the hypoxia-associated ethylene response factor group VII transcription factors of Arabidopsis as substrates of this pathway. Regulation of these proteins by the N-end rule pathway occurs through a characteristic conserved motif at the amino terminus initiating with Met-Cys. Enhanced stability of one of these proteins, HRE2, under low oxygen conditions improves hypoxia survival and reveals a molecular mechanism for oxygen sensing in plants via the evolutionarily conserved N-end rule pathway. SUB1A-1, a major determinant of submergence tolerance in rice3, was shown not to be a substrate for the N-end rule pathway despite containing the N-terminal motif, indicating that it is uncoupled from N-end rule pathway regulation, and that enhanced stability may relate to the superior tolerance of Sub1 rice varieties to multiple abiotic stresses4.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Get just this article for as long as you need it

$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

Figure 1: N-end rule mutants ectopically accumulate anaerobic response mRNAs and are more tolerant to hypoxia.
Figure 2: Group VII ERF transcription factors are substrates for the N-end rule pathway in vitro and in vivo.
Figure 3: HRE proteins are stabilized under low oxygen levels and confer hypoxia tolerance.

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

The microarray data reported in this paper are deposited in Gene Expression Omnibus under accession number GSE29941 and are also tabulated in Supplementary Information.

References

  1. Bailey-Serres, J. & Voesenek, L. A. C. J. Flooding stress: Acclimations and genetic diversity. Annu. Rev. Plant Biol. 59, 313–339 (2008)

    Article  CAS  Google Scholar 

  2. Kaelin, W. G. & Ratcliffe, P. J. Oxygen sensing by metazoans: The central role of the HIF hydroxylase pathway. Mol. Cell 30, 393–402 (2008)

    Article  CAS  Google Scholar 

  3. Xu, K. et al. Sub1A is an ethylene-response-factor-like gene that confers submergence tolerance to rice. Nature 442, 705–708 (2006)

    Article  ADS  CAS  Google Scholar 

  4. Fukao, T., Yeung, E. & Bailey-Serres, J. The submergence tolerance regulator SUB1A mediates crosstalk between submergence and drought tolerance in rice. Plant Cell 23, 412–427 (2011)

    Article  CAS  Google Scholar 

  5. Bachmair, A., Finley, D. & Varshavsky, A. In vivo half-life of a protein is a function of its amino-terminal residue. Science 234, 179–186 (1986)

    Article  ADS  CAS  Google Scholar 

  6. Varshavsky, A. Regulated protein degradation. Trends Biochem. Sci. 30, 283–286 (2005)

    Article  CAS  Google Scholar 

  7. Kwon, Y. T. et al. An essential role of N-terminal arginylation in cardiovascular development. Science 297, 96–99 (2002)

    Article  ADS  CAS  Google Scholar 

  8. Graciet, E., Mesiti, F. & Wellmer, F. Structure and evolutionary conservation of the plant N-end rule pathway. Plant J. 61, 741–751 (2010)

    Article  CAS  Google Scholar 

  9. Graciet, E. & Wellmer, F. The plant N-end rule pathway: structure and functions. Trends Plant Sci. 15, 447–453 (2010)

    Article  CAS  Google Scholar 

  10. Garzón, M. et al. PRT6/At5g02310 encodes an Arabidopsis ubiquitin ligase of the N-end rule pathway with arginine specificity and is not the CER3 locus. FEBS Lett. 581, 3189–3196 (2007)

    Article  Google Scholar 

  11. Tasaki, T. & Kwon, Y. T. The mammalian N-end rule pathway: new insights into its components and physiological roles. Trends Biochem. Sci. 32, 520–528 (2007)

    Article  CAS  Google Scholar 

  12. Holman, T. J. et al. The N-end rule pathway promotes seed germination and establishment through removal of ABA sensitivity in Arabidopsis . Proc. Natl Acad. Sci. USA 106, 4549–4554 (2009)

    Article  ADS  CAS  Google Scholar 

  13. Yoshida, S., Ito, M., Callis, J., Nishida, I. & Watanabe, A. A delayed leaf senescence mutant is defective in arginyl-tRNA: protein arginyltransferase, a component of the N-end rule pathway in Arabidopsis . Plant J. 32, 129–137 (2002)

    Article  CAS  Google Scholar 

  14. Graciet, E. et al. The N-end rule pathway controls multiple functions during Arabidopsis shoot and leaf development. Proc. Natl Acad. Sci. USA 106, 13618–13623 (2009)

    Article  ADS  CAS  Google Scholar 

  15. Mustroph, A. et al. Profiling translatomes of discrete cell populations resolves altered cellular priorities during hypoxia in Arabidopsis . Proc. Natl Acad. Sci. USA 106, 18843–18848 (2009)

    Article  ADS  CAS  Google Scholar 

  16. Chung, H. J. & Ferl, R. J. Arabidopsis alcohol dehydrogenase expression in both shoots and roots is conditioned by root growth environment. Plant Physiol. 121, 429–436 (1999)

    Article  CAS  Google Scholar 

  17. Nakano, T., Suzuki, K., Fujimura, T. & Shinshi, H. Genome-wide analysis of the ERF gene family in Arabidopsis and rice. Plant Physiol. 140, 411–432 (2006)

    Article  CAS  Google Scholar 

  18. Licausi, F. et al. HRE1 and HRE2, two hypoxia-inducible ethylene response factors, affect anaerobic responses in Arabidopsis thaliana . Plant J. 62, 302–315 (2010)

    Article  CAS  Google Scholar 

  19. Hinz, M. et al. Arabidopsis RAP2.2: An ethylene response transcription factor that is important for hypoxia survival. Plant Physiol. 153, 757–772 (2010)

    Article  CAS  Google Scholar 

  20. Papdi, C. et al. Functional identification of Arabidopsis stress regulatory genes using the controlled cDNA overexpression system. Plant Physiol. 147, 528–542 (2008)

    Article  CAS  Google Scholar 

  21. Hattori, Y. et al. The ethylene response factors SNORKEL1 and SNORKEL2 allow rice to adapt to deep water. Nature 460, 1026–1030 (2009)

    Article  ADS  CAS  Google Scholar 

  22. Hu, R. G. et al. The N-end rule pathway as a nitric oxide sensor controlling the levels of multiple regulators. Nature 437, 981–986 (2005)

    Article  ADS  CAS  Google Scholar 

  23. Lee, M. J. et al. RGS4 and RGS5 are in vivo substrates of the N-end rule pathway. Proc. Natl Acad. Sci. USA 102, 15030–15035 (2005)

    Article  ADS  CAS  Google Scholar 

  24. Takahashi, H. et al. A simple and high-sensitivity method for analysis of ubiquitination and polyubiquitination based on wheat cell-free protein synthesis. BMC Plant Biol. 9, 39 (2009)

    Article  Google Scholar 

  25. Suzuki, T. & Varshavsky, A. Degradation signals in the lysine-asparagine sequence space. EMBO J. 18, 6017–6026 (1999)

    Article  CAS  Google Scholar 

  26. Leonard, S. E. & Carroll, K. S. Chemical ‘omics’ approaches for understanding protein cysteine oxidation in biology. Curr. Opin. Chem. Biol. 15, 88–102 (2011)

    Article  CAS  Google Scholar 

  27. Felle, H. H. pH regulation in anoxic plants. Ann. Bot. 96, 519–532 (2005)

    Article  CAS  Google Scholar 

  28. Hu, R. G., Wang, H. Q., Xia, Z. X. & Varshavsky, A. The N-end rule pathway is a sensor of heme. Proc. Natl Acad. Sci. USA 105, 76–81 (2008)

    Article  ADS  CAS  Google Scholar 

  29. Côme, D. & Tissaoui, T. Induction d’une dormance embryonnaire secondaire chez le pommier (Pirus malus L.) par des atmosphères très appauvries en oxygène. Compt. Rendus Acad. Sci. 266, 477–479 (1968)

    Google Scholar 

  30. Branco-Price, C., Kaiser, K. A., Jang, C. J. H., Larive, C. K. & Bailey-Serres, J. Selective mRNA translation coordinates energetic and metabolic adjustments to cellular oxygen deprivation and reoxygenation in Arabidopsis thaliana . Plant J. 56, 743–755 (2008)

    Article  CAS  Google Scholar 

  31. Mustroph, A. et al. Cross-kingdom comparison of transcriptomic adjustments to low-oxygen stress highlights conserved and plant-specific responses. Plant Physiol. 152, 1484–1500 (2010)

    Article  CAS  Google Scholar 

  32. Lucas, M. et al. SHORT-ROOT regulates primary, lateral, and adventitious root development in Arabidopsis . Plant Physiol. 155, 384–398 (2011)

    Article  CAS  Google Scholar 

  33. Dubin, M. J., Bowler, C. & Benvenuto, G. A modified Gateway cloning strategy for overexpressing tagged proteins in plants. Plant Methods 4, 3 (2008)

    Article  Google Scholar 

  34. Swarup, R. et al. Root gravitropism requires lateral root cap and epidermal cells for transport and response to a mobile auxin signal. Nature Cell Biol. 7, 1057–1065 (2005)

    Article  CAS  Google Scholar 

  35. Martinez-Garcia, J. F., Monte, E. & Quail, P. H. A simple, rapid and quantitative method for preparing Arabidopsis protein extracts for immunoblot analysis. Plant J. 20, 251–257 (1999)

    Article  CAS  Google Scholar 

  36. Williams, A. J., Werner-Fraczek, J., Chang, I. F. & Bailey-Serres, J. Regulated phosphorylation of 40S ribosomal protein S6 in root tips of maize. Plant Physiol. 132, 2086–2097 (2003)

    Article  CAS  Google Scholar 

  37. Thompson, J. D., Higgins, D. G. & Gibson, T. J. Clustal-W - Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22, 4673–4680 (1994)

    Article  CAS  Google Scholar 

  38. Horan, K. et al. Annotating genes of known and unknown function by large-scale coexpression analysis. Plant Physiol. 147, 41–57 (2008)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

M.J.H., D.J.G., S.G. and C.S.C. were supported by BBSRC grant BB/G010595/1; G.W.B. by a Marie Curie International Incoming Fellowship; N.M.I. by a MARA PhD fellowship from the Malaysian government; S.C.L., T.F. and J.B.-S. by grants NSF IOS-0750811 and NIFA 2008-35100-04528. We thank S. Liddell. Rothamsted Research receives grant-aided support from the BBSRC.

Author information

Author notes

  1. Daniel J. Gibbs and Seung Cho Lee: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Plant and Crop Sciences, School of Biosciences and Centre for Plant Integrative Biology, University of Nottingham, Loughborough LE12 5RD, UK

    Daniel J. Gibbs, Nurulhikma Md Isa, Silvia Gramuglia, George W. Bassel, Cristina Sousa Correia & Michael J. Holdsworth

  2. Center for Plant Cell Biology and Department of Botany and Plant Sciences, University of California, Riverside, 92521, California, USA

    Seung Cho Lee, Takeshi Fukao & Julia Bailey-Serres

  3. UPMC Univ Paris 06, UR5-EAC 7180 CNRS, Boîte courrier 156, 4 place Jussieu, F-75005 Paris, France

    Françoise Corbineau

  4. Biological Chemistry Department, Rothamsted Research, Harpenden AL5 2JQ, UK

    Frederica L. Theodoulou

Authors
  1. Daniel J. Gibbs
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Seung Cho Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nurulhikma Md Isa
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Silvia Gramuglia
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Takeshi Fukao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. George W. Bassel
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Cristina Sousa Correia
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Françoise Corbineau
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Frederica L. Theodoulou
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Julia Bailey-Serres
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Michael J. Holdsworth
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

D.J.G., M.J.H., J.B.-S., F.C. and F.L.T. conceived and designed experiments. D.J.G., S.C.L., N.M.I., S.G., C.S.C., G.W.B., T.F. and F.C. performed the experiments. D.J.G., S.C.L., N.M.I., S.G., C.S.C., G.W.B., T.F., F.C. M.J.H., J.B-S. and F.L.T. analysed the data. M.J.H., D.J.G. and J.B.-S. wrote the manuscript.

Corresponding authors

Correspondence to Julia Bailey-Serres or Michael J. Holdsworth.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

The file contains Supplementary Figures 1-5 with legends. (PDF 718 kb)

Supplementary Table 1

This file contains microarray data comparing gene expression in Col-0 (WT), prt6-1 and ate1-2 ate2-1 mutants. (XLS 310 kb)

Supplementary Table 2

This file contains a list of oligonucleotide primers used in this study. (XLS 14 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Gibbs, D., Lee, S., Md Isa, N. et al. Homeostatic response to hypoxia is regulated by the N-end rule pathway in plants. Nature 479, 415–418 (2011). https://doi.org/10.1038/nature10534

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature10534

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing