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Published on: December 14, 2017
STARD3 regulates lysosome positioning and contacts via a GSK3-controlled phosphorylation switch
Julie Eichler1,2,3,4, Corinne Wendling1,2,3,4, Sophie Huver1,2,3,4
1Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Illkirch, France.
Glycogen synthase kinase 3 (GSK3) phosphorylates STARD3, activating its role in forming membrane contact sites (MCS) between late endosomes/lysosomes and the endoplasmic reticulum. This phosphorylation is crucial for organelle communication and endosome function.
Area of Science:
- Cell Biology
- Molecular Biology
- Organelle Biology
Background:
- Membrane contact sites (MCS) are crucial for cellular homeostasis, facilitating lipid exchange and organelle positioning.
- STARD3 is a cholesterol transfer protein that forms contacts between late endosomes/lysosomes (LE/Lys) and the endoplasmic reticulum (ER) via its Phospho-FFAT motif.
- The interaction of STARD3's motif with ER-resident VAPs is regulated by phosphorylation.
Purpose of the Study:
- To identify the kinases responsible for phosphorylating STARD3.
- To elucidate the role of STARD3 phosphorylation in regulating ER-LE/Lys membrane contact sites.
- To investigate additional functions of STARD3 in endosome biology.
Main Methods:
- Kinase assays to identify STARD3-phosphorylating enzymes.
- Phosphorylation site mapping within the STARD3 Phospho-FFAT motif.
- Biochemical and cell-based assays to assess STARD3 tethering activity and ER-LE/Lys contact formation.
- Analysis of LE/Lys homotypic interactions.
Main Results:
- GSK3α and GSK3β were identified as the kinases that phosphorylate STARD3 at serine 209 within its Phospho-FFAT motif.
- Phosphorylation of STARD3 at serine 209 is necessary and sufficient for activating its tethering function, promoting ER-LE/Lys contacts.
- STARD3 also mediates LE/Lys homotypic interactions when ER-LE/Lys tethering is impaired, indicating a dual role in endosome organization.
Conclusions:
- GSK3 directly regulates membrane contact site formation through STARD3 phosphorylation.
- This study expands the understanding of the molecular mechanisms governing inter-organelle communication and endosome function.
- STARD3 acts as a key regulator of both ER-LE/Lys contacts and LE/Lys homotypic interactions.
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