LRRK2 mediated Rab8a phosphorylation promotes lipid storage
Abstract Background Several mutations in leucine rich repeat kinase 2 (LRRK2) gene have been associated with pathogenesis of Parkinson’s disease (PD), a neurodegenerative disorder marked by resting tremors, and rigidity, leading to Postural instability. It has been revealed that mutations that lead...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-02-01
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| Series: | Lipids in Health and Disease |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12944-018-0684-x |
| _version_ | 1818840199235371008 |
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| author | Miao Yu Muhammad Arshad Wenmin Wang Dongyu Zhao Li Xu Linkang Zhou |
| author_facet | Miao Yu Muhammad Arshad Wenmin Wang Dongyu Zhao Li Xu Linkang Zhou |
| author_sort | Miao Yu |
| collection | DOAJ |
| description | Abstract Background Several mutations in leucine rich repeat kinase 2 (LRRK2) gene have been associated with pathogenesis of Parkinson’s disease (PD), a neurodegenerative disorder marked by resting tremors, and rigidity, leading to Postural instability. It has been revealed that mutations that lead to an increase of kinase activity of LRRK2 protein are significantly associated with PD pathogenesis. Recent studies have shown that some Rab GTPases, especially Rab8, serve as substrates of LRRK2 and undergo phosphorylation in its switch II domain upon interaction. Current study was performed in order to find out the effects of the phosphorylation of Rab8 and its mutants on lipid metabolism and lipid droplets growth. Methods The phosphorylation status of Rab8a was checked by phos-tag gel. Point mutant construct were generated to investigate the function of Rab8a. 3T3L1 cells were transfected with indicated plasmids and the lipid droplets were stained with Bodipy. Fluorescent microscopy experiments were performed to examine the sizes of lipid droplets. The interactions between Rab8a and Optineurin were determined by immunoprecipitation and western blot. Results Our assays demonstrated that Rab8a was phosphorylated by mutated LRRK2 that exhibits high kinase activity. Phosphorylation of Rab8a on amino acid residue T72 promoted the formation of large lipid droplets. T72D mutant of Rab8a had higher activity to promote the formation of large lipid droplets compared with wild type Rab8a, with increase in average diameter of lipid droplets from 2.10 μm to 2.46 μm. Moreover, phosphorylation of Rab8a weakened the interaction with its effector Optineurin. Conclusions Y1699C mutated LRRK2 was able to phosphorylate Rab8a and phosphorylation of Rab8a on site 72 plays important role in the fusion and enlargement of lipid droplets. Taken together, our study suggests an indirect relationship between enhanced lipid storage capacity and PD pathogenesis. |
| first_indexed | 2024-12-19T04:06:23Z |
| format | Article |
| id | doaj.art-eecbb85ad5784d78ba0a4ff1b33f97b5 |
| institution | Directory Open Access Journal |
| issn | 1476-511X |
| language | English |
| last_indexed | 2024-12-19T04:06:23Z |
| publishDate | 2018-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Lipids in Health and Disease |
| spelling | doaj.art-eecbb85ad5784d78ba0a4ff1b33f97b52022-12-21T20:36:32ZengBMCLipids in Health and Disease1476-511X2018-02-011711810.1186/s12944-018-0684-xLRRK2 mediated Rab8a phosphorylation promotes lipid storageMiao Yu0Muhammad Arshad1Wenmin Wang2Dongyu Zhao3Li Xu4Linkang Zhou5MOE Key Laboratory of Bioinformatics and Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua UniversityDepartment of Bioinformatics and Biotechnology, International Islamic UniversityMOE Key Laboratory of Bioinformatics and Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua UniversityMOE Key Laboratory of Bioinformatics and Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua UniversityMOE Key Laboratory of Bioinformatics and Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua UniversityMOE Key Laboratory of Bioinformatics and Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua UniversityAbstract Background Several mutations in leucine rich repeat kinase 2 (LRRK2) gene have been associated with pathogenesis of Parkinson’s disease (PD), a neurodegenerative disorder marked by resting tremors, and rigidity, leading to Postural instability. It has been revealed that mutations that lead to an increase of kinase activity of LRRK2 protein are significantly associated with PD pathogenesis. Recent studies have shown that some Rab GTPases, especially Rab8, serve as substrates of LRRK2 and undergo phosphorylation in its switch II domain upon interaction. Current study was performed in order to find out the effects of the phosphorylation of Rab8 and its mutants on lipid metabolism and lipid droplets growth. Methods The phosphorylation status of Rab8a was checked by phos-tag gel. Point mutant construct were generated to investigate the function of Rab8a. 3T3L1 cells were transfected with indicated plasmids and the lipid droplets were stained with Bodipy. Fluorescent microscopy experiments were performed to examine the sizes of lipid droplets. The interactions between Rab8a and Optineurin were determined by immunoprecipitation and western blot. Results Our assays demonstrated that Rab8a was phosphorylated by mutated LRRK2 that exhibits high kinase activity. Phosphorylation of Rab8a on amino acid residue T72 promoted the formation of large lipid droplets. T72D mutant of Rab8a had higher activity to promote the formation of large lipid droplets compared with wild type Rab8a, with increase in average diameter of lipid droplets from 2.10 μm to 2.46 μm. Moreover, phosphorylation of Rab8a weakened the interaction with its effector Optineurin. Conclusions Y1699C mutated LRRK2 was able to phosphorylate Rab8a and phosphorylation of Rab8a on site 72 plays important role in the fusion and enlargement of lipid droplets. Taken together, our study suggests an indirect relationship between enhanced lipid storage capacity and PD pathogenesis.http://link.springer.com/article/10.1186/s12944-018-0684-xLRRK2Rab8aLipid dropletsLipid storageParkinson’s disease |
| spellingShingle | Miao Yu Muhammad Arshad Wenmin Wang Dongyu Zhao Li Xu Linkang Zhou LRRK2 mediated Rab8a phosphorylation promotes lipid storage Lipids in Health and Disease LRRK2 Rab8a Lipid droplets Lipid storage Parkinson’s disease |
| title | LRRK2 mediated Rab8a phosphorylation promotes lipid storage |
| title_full | LRRK2 mediated Rab8a phosphorylation promotes lipid storage |
| title_fullStr | LRRK2 mediated Rab8a phosphorylation promotes lipid storage |
| title_full_unstemmed | LRRK2 mediated Rab8a phosphorylation promotes lipid storage |
| title_short | LRRK2 mediated Rab8a phosphorylation promotes lipid storage |
| title_sort | lrrk2 mediated rab8a phosphorylation promotes lipid storage |
| topic | LRRK2 Rab8a Lipid droplets Lipid storage Parkinson’s disease |
| url | http://link.springer.com/article/10.1186/s12944-018-0684-x |
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