Important for microtubules and nerve growth
This study shines light on how levels of the Spastin protein are regulated in HSP, and the effects this has on microtubule dynamics and neurite outgrowth.
Abstract
Spastin is one of the proteins which lead to hereditary spastic paraplegia (HSP), whose dysfunction towards microtubule severing and membrane transporting is critically important. The present study is to elucidate the mechanisms of the protein stability regulation of spastin.
The ubiquitin encoding plasmids are transfected into COS-7 cells with different fusion tags including Green Fluorescent Protein (GFP), mCherry and Flag. The expression level of spastin was detected, microtubule severing activity and neurite outgrowth were quantified. The data showed that ubiquitin overexpression significantly induced the decreased expression of spastin, suppressed the activity of microtubule severing in COS-7 cells and inhibited the promoting effect on neurite outgrowth in cultured hippocampal neurons. Furthermore, when modulating the overexpression experiments of ubiquitin, it was found that relatively small tags like Flag, but not large tags such as GFP or mCherry fused with ubiquitin, retained the activity on spastin stability.
The present study investigated the effects of small/large tags addition to ubiquitin and the novel mechanisms of post-transcriptional modifications of spastin on regulating neurite outgrowth, in the attempt to experimentally elucidate the mechanisms that control the level or stability of spastin in hereditary spastic paraplegia.
SOURCE: Eur J Histochem. 2021 Dec 7;65(4):3352. doi: 10.4081/ejh.2021.3352. PMID: 34873900
Different fusion tags affect the activity of ubiquitin overexpression on spastin protein stability
Jianyu Zou 1 , Zhenbin Cai 2 , Zhi Liang 3 , Yaozhong Liang 4 , Guowei Zhang 5 , Jie Yang 6 , Yunlong Zhang 7 , Hongsheng Lin 8 , Minghui Tan 7
1. Department of Orthopaedics, The First Affiliated Hospital of Jinan University, Guangzhou. [email protected].
2. Department of Orthopaedics, The First Affiliated Hospital of Jinan University, Guangzhou. [email protected].
3. Department of Orthopaedics, The Eighth Affiliated Hospital of Sun Yat-sen University, Shenzhen. [email protected].
4. Department of Orthopaedics, The First Affiliated Hospital of Jinan University, Guangzhou. [email protected].
5. Department of Orthopaedics, The First Affiliated Hospital of Jinan University, Guangzhou. [email protected].
6. Department of Orthopaedics, The First Affiliated Hospital of Jinan University, Guangzhou. [email protected].
7. Department of Orthopaedics, The First Affiliated Hospital of Jinan University, Guangzhou. [email protected].
8. Department of Orthopaedics, The First Affiliated Hospital of Jinan University, Guangzhou. [email protected].