Identification of downstream components of ubiquitin-conjugating enzyme PHOSPHATE2 by quantitative membrane proteomics in Arabidopsis roots

Teng Kuei Huang, Chia Li Han, Shu I. Lin, Yu Ju Chen, Yi Chuan Tsai, Yet Ran Chen, June Wei Chen, Wei Yi Lin, Pei Mien Chen, Tzu Yin Liu, Ying Shin Chen, Ching Mei Sun, Tzyy Jen Chiou

研究成果: 雜誌貢獻文章

125 引文 斯高帕斯(Scopus)

摘要

MicroRNA399-mediated regulation of the ubiquitin-conjugating enzyme UBC24/PHOSPHATE2 (PHO2) is crucial for Pi acquisition and translocation in plants. Because of a potential role for PHO2 in protein degradation and its association with membranes, an iTRAQ (for isobaric tags for relative and absolute quantitation)- based quantitative membrane proteomic method was employed to search for components downstream of PHO2. A total of 7491 proteins were identified from Arabidopsis thaliana roots by mass spectrometry, 35.2% of which were predicted to contain at least one transmembrane helix. Among the quantifiable proteins, five were significantly differentially expressed between the wild type and pho2 mutant under two growth conditions. Using immunoblot analysis, we validated the upregulation of several members in PHOSPHATE TRANSPORTER1 (PHT1) family and PHOSPHATE TRANSPORTER TRAFFIC FACILITATOR1 (PHF1) in pho2 and demonstrated that PHO2 mediates the degradation of PHT1 proteins. Genetic evidence that loss of PHF1 or PHT1;1 alleviated Pi toxicity in pho2 further suggests that they play roles as downstream components of PHO2. Moreover, we showed that PHO2 interacts with PHT1s in the postendoplasmic reticulum compartments and mediates the ubiquitination of endomembrane-localized PHT1;1. This study not only uncovers a mechanism by which PHO2 modulates Pi acquisition by regulating the abundance of PHT1s in the secretory pathway destined for plasma membranes, but also provides a database of the membrane proteome that will be widely applicable in root biology research.

原文英語
頁(從 - 到)4044-4060
頁數17
期刊Plant Cell
25
發行號10
DOIs
出版狀態已發佈 - 十月 2013
對外發佈Yes

ASJC Scopus subject areas

  • Plant Science
  • Cell Biology
  • Medicine(all)

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  • 引用此

    Huang, T. K., Han, C. L., Lin, S. I., Chen, Y. J., Tsai, Y. C., Chen, Y. R., Chen, J. W., Lin, W. Y., Chen, P. M., Liu, T. Y., Chen, Y. S., Sun, C. M., & Chiou, T. J. (2013). Identification of downstream components of ubiquitin-conjugating enzyme PHOSPHATE2 by quantitative membrane proteomics in Arabidopsis roots. Plant Cell, 25(10), 4044-4060. https://doi.org/10.1105/tpc.113.115998