Molecular characterization of SOF10 as a FREE1 suppressor in Arabidopsis
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AbstractMolecular characterization of SOF10 as a FREE1 suppressor in Arabidopsis
Ying ZHU1*, Jinbo SHEN1, Qiong ZHAO1 and Liwen JIANG1,2
1 School of Life Sciences, Centre for Cell & Developmental Biology and State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China
2 The Chinese University of Hong Kong Shenzhen Research Institute, Shenzhen 518057, China

All eukaryotic cells contain several functionally distinct membrane-enclosed organelles in their endomembrane system, including the endoplasmic reticulum (ER), Golgi apparatus, trans-Golgi network (TGN), endosomes, prevacuolar compartment (PVC) or multivesicular body (MVB) and vacuole/lysosome. The endosomal sorting complex required for transport (ESCRT) machinery mediates the formation of intraluminal vesicles (ILVs) in MVBs as well as the sorting of the ubiquitinated cargoes into the internal vesicles. We have recently characterized a plant unique ESCRT key component termed FYVE domain protein required for endosomal sorting 1 (FREE1) and demonstrated that functional loss of FREE1 in Arabidopsis showed seedling lethal phenotype, fragmented vacuoles, MVB ILVs biogenesis defection, and accumulated autophagosomes. To further illustrate the underlying mechanisms of FREE1 multiple functions in plants, a forward genetic screen was performed for mutants that suppressed the seedling lethal phenotype of FREE1-RNAi transgenic plants and the obtained mutants are termed suppressors of free1 (sof). Next generation sequencing analysis identified two sof lines (sof10 and sof641) mapped to the same causal gene SOF10A. The sof10 possesses the mutation which results in a premature stop codon, while the sof641 produced a mis-sense mutation. Both sof10 and sof641 seedlings exhibited reduced FREE1 protein level, normal vacuole and MVB morphology, and normal membrane protein degradation. Preliminary subcellular localization results of SOF10A showed cytosol pattern with some punctate dots. In Arabidopsis, SOF10A has a close homolog termed SOF10B. Either SOF10A or SOF10B T-DNA insertional mutants can restore the lethal phenotype of FREE1-RNAi mutants, whereas the double homozygous T-DNA insertional mutant of SOF10A and SOF10B is lethal. Further characterization of SOF10A and SOF10B combined with cell and genetic approaches will illustrate the underlying mechanisms of SOF10 as FREE1 suppressor and its function in membrane protein trafficking and organelle biogenesis in plants. Here we will present an update on this study. Supported by Research Grants Council of Hong Kong and G-CUHK403/17.

References
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Zhao, Q.; Gao, C.; Lee, P; Liu, L.; Li, S.; Hu, T.; Shen, J.; Pan, S.; Ye, H.; Chen, Y.; Cao, W.; Cui, Y.; Zeng, P.; Yu, S.; Gao, Y.; Chen, L.; Mo, B.; Liu, X.; Xiao, S.; Zhao, Y.; Zhong, S.; Chen, X.; Jiang, L. 2015. Fast-suppressor Screening for New Components in Protein Trafficking, Organelle Biogenesis and Silencing Pathway in Arabidopsis thaliana using DEX-Inducible FREE1-RNAi Plants. Journal of Genetics and Genomics 42(6): 319-330.

The 21st Meeting of the European Network for Plant Endomembrane Research. September 4th- 7th, 2018. Vienna, Austria
All Author(s) ListYing ZHU, Jinbo SHEN, Qiong ZHAO, Liwen JIANG
Name of ConferenceThe 21st Meeting of the European Network for Plant Endomembrane Research
Start Date of Conference04/09/2018
End Date of Conference07/09/2018
Place of ConferenceVienna
Country/Region of ConferenceAustria
Year2018
LanguagesEnglish-United States

Last updated on 2018-23-10 at 12:10