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

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)4044-4060
Number of pages17
JournalPlant Cell
Volume25
Issue number10
DOIs
Publication statusPublished - Oct 2013
Externally publishedYes

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Ubiquitin-Conjugating Enzymes
ubiquitin-protein ligase
Arabidopsis
Phosphate Transport Proteins
Proteomics
proteomics
traffic
Membranes
transporters
phosphates
Reticulum
Proteins
reticulum
proteins
Secretory Pathway
Ubiquitination
Proteome
protein degradation
proteome
Proteolysis

ASJC Scopus subject areas

  • Plant Science
  • Cell Biology
  • Medicine(all)

Cite this

Identification of downstream components of ubiquitin-conjugating enzyme PHOSPHATE2 by quantitative membrane proteomics in Arabidopsis roots. / Huang, Teng Kuei; Han, Chia Li; Lin, Shu I.; Chen, Yu Ju; Tsai, Yi Chuan; Chen, Yet Ran; Chen, June Wei; Lin, Wei Yi; Chen, Pei Mien; Liu, Tzu Yin; Chen, Ying Shin; Sun, Ching Mei; Chiou, Tzyy Jen.

In: Plant Cell, Vol. 25, No. 10, 10.2013, p. 4044-4060.

Research output: Contribution to journalArticle

Huang, TK, Han, CL, Lin, SI, Chen, YJ, Tsai, YC, Chen, YR, Chen, JW, Lin, WY, Chen, PM, Liu, TY, Chen, YS, Sun, CM & Chiou, TJ 2013, 'Identification of downstream components of ubiquitin-conjugating enzyme PHOSPHATE2 by quantitative membrane proteomics in Arabidopsis roots', Plant Cell, vol. 25, no. 10, pp. 4044-4060. https://doi.org/10.1105/tpc.113.115998
Huang, Teng Kuei ; Han, Chia Li ; Lin, Shu I. ; Chen, Yu Ju ; Tsai, Yi Chuan ; Chen, Yet Ran ; Chen, June Wei ; Lin, Wei Yi ; Chen, Pei Mien ; Liu, Tzu Yin ; Chen, Ying Shin ; Sun, Ching Mei ; Chiou, Tzyy Jen. / Identification of downstream components of ubiquitin-conjugating enzyme PHOSPHATE2 by quantitative membrane proteomics in Arabidopsis roots. In: Plant Cell. 2013 ; Vol. 25, No. 10. pp. 4044-4060.
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AU - Chen, Yu Ju

AU - Tsai, Yi Chuan

AU - Chen, Yet Ran

AU - Chen, June Wei

AU - Lin, Wei Yi

AU - Chen, Pei Mien

AU - Liu, Tzu Yin

AU - Chen, Ying Shin

AU - Sun, Ching Mei

AU - Chiou, Tzyy Jen

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