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Identification of three hydroxyproline O-arabinosyltransferases in Arabidopsis thaliana

Nature Chemical Biology volume 9, pages 726–730 (2013)Cite this article

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Abstract

Hydroxyproline (Hyp) O-arabinosylation is a post-translational modification that is prominent in extracellular glycoproteins in plants. Hyp O-arabinosylation is generally found in these glycoproteins in the form of linear oligoarabinoside chains and has a key role in their function by contributing to conformational stability. However, Hyp O-arabinosyltransferase (HPAT), a key enzyme that catalyzes the transfer of the L-arabinose to the hydroxyl group of Hyp residues, has remained undiscovered. Here, we purified and identified Arabidopsis HPAT as a Golgi-localized transmembrane protein that is structurally similar to the glycosyltransferase GT8 family. Loss-of-function mutations in HPAT-encoding genes cause pleiotropic phenotypes that include enhanced hypocotyl elongation, defects in cell wall thickening, early flowering, early senescence and impaired pollen tube growth. Our results indicate essential roles of Hyp O-arabinosylation in both vegetative and reproductive growth in plants.

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Figure 1: Purification and identification of Arabidopsis HPAT.
Figure 2: Phenotypes of loss-of-function mutants of HPAT1, HPAT2 and HPAT3.
Figure 3: Decrease in Hyp O-arabinosylation of EXT3 in hpat mutants.
Figure 4: Decrease in Hyp O-arabinosylation of CLE2 in hpat mutants.

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Acknowledgements

We thank the National Institute for Basic Biology Functional Genomics Facility for MALDI-TOF MS analysis and T. Ueda (University of Tokyo) for providing the mRFP-SYP31 expression vector. This research was supported by the Funding Program for Next Generation World-Leading Researchers from the Japan Society for the Promotion of Science (no. GS025) and Grants-in-Aid for Scientific Research (S) from the Ministry of Education, Culture, Sports, Science and Technology (no. 25221105).

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Authors and Affiliations

  1. National Institute for Basic Biology, Okazaki, Japan

    Mari Ogawa-Ohnishi & Yoshikatsu Matsubayashi

  2. Graduate School of Bio-Agricultural Sciences, Nagoya University, Nagoya, Japan

    Wataru Matsushita

Authors
  1. Mari Ogawa-Ohnishi
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  2. Wataru Matsushita
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  3. Yoshikatsu Matsubayashi
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Contributions

M.O.-O. and Y.M. designed experiments. M.O.-O., W.M. and Y.M. performed and analyzed experiments. M.O.-O. and Y.M. wrote the manuscript. Y.M. supervised the project.

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Correspondence to Yoshikatsu Matsubayashi.

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The authors declare no competing financial interests.

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Supplementary Results, Supplementary Tables 1–3 and Supplementary Figures 1–10. (PDF 8342 kb)

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Ogawa-Ohnishi, M., Matsushita, W. & Matsubayashi, Y. Identification of three hydroxyproline O-arabinosyltransferases in Arabidopsis thaliana. Nat Chem Biol 9, 726–730 (2013). https://doi.org/10.1038/nchembio.1351

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