Arabidopsis Qa-SNARE SYP2 proteins localized to different subcellular regions function redundantly in vacuolar protein sorting and plant development

Makoto Shirakawa, Haruko Ueda, Tomoo Shimada, Yasuko Koumoto, Takashi L. Shimada, Maki Kondo, Taku Takahashi, Yudai Okuyama, Mikio Nishimura, Ikuko Hara-Nishimura

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Abstract

SYP2 proteins are a sub-family of Qa-SNAREs (soluble N-ethylmaleimide- sensitive factor attachment protein receptors) that may be responsible for protein trafficking between pre-vacuolar compartments (PVC) and vacuoles. Arabidopsis thaliana SYP22/VAM3/SGR3 and SYP21/PEP12 proteins function independently, but are both reported to be essential for male gametophytic viability. Here, we systematically examined the redundancy of three SYP2 paralogs (i.e. SYP21, 22 and 23) using a Col-0 ecotype harboring a SYP2 paralog (SYP23/PLP) that lacked a transmembrane domain. Surprisingly, no visible phenotypes were observed, even in the double knockout syp21/pep12 syp23/plp. Deficiency of either SYP21/PEP12 or SYP23/PLP in the syp22 background resulted in a defect in vacuolar protein sorting, characterized by abnormal accumulation of protein precursors in seeds. SYP21/PEP12 knockdown enhanced the syp22 phenotype (i.e. semi-dwarfism, poor leaf vein development and abnormal development of myrosin cells), and additional knockout of SYP23/PLP further aggravated the phenotype. A GFP-SYP23/PLP fusion localized to the cytosol, but not to the PVC or vacuolar membrane, where SYP21/PEP12 or SYP22/VAM3, respectively, were localized. Immunoprecipitation analysis showed that SYP23/PLP interacted with the vacuolar Qb- and Qc-SNAREs, VTI11 and SYP5, respectively, suggesting that SYP23/PLP is able to form a SNARE complex anchoring the membrane. Unexpectedly, we found that expression of multiple copies of a genomic fragment of SYP23/PLP suppressed the abnormal syp22-3 phenotype. Thus, SYP2 proteins, including cytosolic SYP23/PLP, appear to function redundantly in vacuolar trafficking and plant development.

Original languageEnglish
Pages (from-to)924-935
Number of pages12
JournalPlant Journal
Volume64
Issue number6
DOIs
Publication statusPublished - Dec 2010

Fingerprint

Arabidopsis Proteins
Qa-SNARE Proteins
Plant Development
protein transport
Protein Transport
plant development
Arabidopsis
Phenotype
SNARE Proteins
Qc-SNARE Proteins
vacuoles
Qb-SNARE Proteins
phenotype
proteins
Ecotype
Dwarfism
Proteins
Protein Precursors
Membranes
Vacuoles

Keywords

  • Arabidopsis thaliana
  • myrosin cell
  • plant development
  • SNARE
  • vacuolar protein sorting
  • vein patterning

ASJC Scopus subject areas

  • Plant Science
  • Cell Biology
  • Genetics

Cite this

Arabidopsis Qa-SNARE SYP2 proteins localized to different subcellular regions function redundantly in vacuolar protein sorting and plant development. / Shirakawa, Makoto; Ueda, Haruko; Shimada, Tomoo; Koumoto, Yasuko; Shimada, Takashi L.; Kondo, Maki; Takahashi, Taku; Okuyama, Yudai; Nishimura, Mikio; Hara-Nishimura, Ikuko.

In: Plant Journal, Vol. 64, No. 6, 12.2010, p. 924-935.

Research output: Contribution to journalArticle

Shirakawa, M, Ueda, H, Shimada, T, Koumoto, Y, Shimada, TL, Kondo, M, Takahashi, T, Okuyama, Y, Nishimura, M & Hara-Nishimura, I 2010, 'Arabidopsis Qa-SNARE SYP2 proteins localized to different subcellular regions function redundantly in vacuolar protein sorting and plant development', Plant Journal, vol. 64, no. 6, pp. 924-935. https://doi.org/10.1111/j.1365-313X.2010.04394.x
Shirakawa, Makoto ; Ueda, Haruko ; Shimada, Tomoo ; Koumoto, Yasuko ; Shimada, Takashi L. ; Kondo, Maki ; Takahashi, Taku ; Okuyama, Yudai ; Nishimura, Mikio ; Hara-Nishimura, Ikuko. / Arabidopsis Qa-SNARE SYP2 proteins localized to different subcellular regions function redundantly in vacuolar protein sorting and plant development. In: Plant Journal. 2010 ; Vol. 64, No. 6. pp. 924-935.
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