Meihen Sun; Xu Han; Fei Chang; Hongfei Xu; Lesley Colgan; Yong-Jian Liu

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Meihen Sun, Xu Han, Fei Chang, Hongfei Xu, Lesley Colgan, Yong-Jian Liu. Regulatory role of sorting nexin 5 in protein stability and vesicular targeting of vesicular acetylcholine transporter to synaptic vesicle-like vesicles in PC12 Cells.[J]. The Journal of Biomedical Research.

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Regulatory role of sorting nexin 5 in protein stability and vesicular targeting of vesicular acetylcholine transporter to synaptic vesicle-like vesicles in PC12 Cells.

Meihen Sun^{1, #},
Xu Han^{1, #},
Fei Chang^{2, #},
Hongfei Xu³,
Lesley Colgan^{4
,
,},
Yong-Jian Liu^{1
,
,}

1.
Jiangsu Key Laboratory of Xenotransplantation, and Department of Medical Genetics Nanjing Medical University, Nanjing 211166, China
2.
Neuroscience program, University of Utah School of Medicine, Utah, 84112 USA
3.
Department of Neurology, UCSF School of Medicine, California, 94143 USA
4.
Max Planck Florida Institute for Neuroscience, Florida 33458, USA

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Corresponding author: Lesley Colgan, Max Planck Florida Institute for Neuroscience, Jupiter, Florida 33458, USA, Tel: 01-561-972-9210, E-mail: lesley.colgan@nufi.org; Yong-Jian Liu, Jiangsu Key Laboratory of Xenotransplantation, and Department of Medical Genetics, Nanjing Medical University, Nanjing 211166, China, Tel: 86-25-86869442; E-mail: young.liu78@gmail.com

Abstract

Abstract

Accurate targeting of vesicular acetylcholine transporter (VAChT) to synaptic vesicles (SVs) is indispensable for efficient cholinergic transmission. Previous studies have suggested that the dileucine motif within the C-terminus of the transporter is sufficient for its targeting to SVs. However, the cytosolic mechanism underlying specific regulation of VAChT trafficking and targeting to SVs is still unclear. Here we use the C-terminus of VAChT as bait in a yeast-two-hybrid screen to identify sorting nexin 5 (SNX5) as its novel interacting protein. SNX5 is detected in the SVs enriched LP2 subcellular fraction of rat brain homogenate and shows strong colocalization with VAChT in both brain sections and PC12 cells. Binding assays suggest that the C-terminal domain of VAChT can interact with both BAR and PX domain of SNX5. Depletion of SNX5 enhances the degradation of VAChT and the process is mediated through the lysosomal pathway. More importantly, we find that, in PC12 cells, the depletion of SNX5 expression significantly decreases the synaptic vesicle-like vesicles (SVLVs) localization of VAChT. Therefore, the results suggest that SNX5 is a novel regulator for both stability and SV targeting of VAChT.
- retrograde trafficking,
- vesicular targeting,
- synaptic vesicle-like vesicles,
- synaptic vesicles,
- PC12

#These authors contributed equally to this work.

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References(38)

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