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

• Published in: The Plant journal : for cell and molecular biology Vol. 64; no. 6; pp. 924 - 935
• Main Authors: Shirakawa, Makoto, Ueda, Haruko, Shimada, Tomoo, Koumoto, Yasuko, Shimada, Takashi L., Kondo, Maki, Takahashi, Taku, Okuyama, Yudai, Nishimura, Mikio, Hara‐Nishimura, Ikuko
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.12.2010; Blackwell
• Subjects: Arabidopsis - genetics; Arabidopsis - growth & development; Arabidopsis - metabolism; Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Arabidopsis thaliana; Biological and medical sciences; Cell Differentiation; Cell differentiation, maturation, development, hematopoiesis; Cell physiology; Flowers & plants; Fundamental and applied biological sciences. Psychology; Gene Knockout Techniques; Gene loci; Molecular and cellular biology; Mutation; myrosin cell; Phenotype; plant development; Plant physiology and development; Plants, Genetically Modified - genetics; Plants, Genetically Modified - growth & development; Plants, Genetically Modified - metabolism; Protein Transport; Proteins; Qa-SNARE Proteins - genetics; Qa-SNARE Proteins - metabolism; Secretion. Exocytosis; SNARE; vacuolar protein sorting; Vacuoles - metabolism; vein patterning; Patterning; Vegetals; myrosin cell; Exocytosis; SNARE; Arabidopsis thaliana; vacuolar protein sorting; plant development; Plant; vein patterning; Cruciferae; Dicotyledones; Angiospermae; Development; Protein sorting; Spermatophyta; Vein; Cell
• ISSN: 0960-7412; 1365-313X
• DOI: 10.1111/j.1365-313X.2010.04394.x

Summary 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.