Tankyrase regulates epithelial lumen formation via suppression of Rab11 GEFs
Arun A Chandrakumar1,2, Étienne Coyaud2, Christopher B Marshall2
1Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.
Tankyrase suppresses SH3BP5 and SH3BP5L, inhibiting Rab11a activation and cyst lumen formation. RNF146 reverses this, restoring epithelial polarity and lumen development.
Area of Science:
- Cell Biology
- Molecular Biology
- Epithelial Biology
Background:
- Rab11 GTPases are essential for cell division, cilia formation, and creating lumens within tissues.
- Rab11a specifically facilitates the transport of podocalyxin (PODXL) to the apical surface during epithelial lumen formation.
- SH3BP5 and SH3BP5L act as guanine nucleotide exchange factors (GEFs) crucial for activating Rab11.
Purpose of the Study:
- To investigate the role of SH3BP5 and SH3BP5L in Rab11a activation and epithelial cyst lumen formation.
- To identify upstream regulators of SH3BP5 and SH3BP5L using proteomic approaches.
- To elucidate the signaling pathway governing epithelial polarity and lumen development.
Main Methods:
- Proximity-dependent biotin identification (BioID) interaction proteomics to identify protein interactors.
- Biochemical assays to assess GEF activity and protein-protein interactions.
- Cell culture models of epithelial cyst formation to study lumen development.
Main Results:
- SH3BP5 and SH3BP5L were identified as substrates of Tankyrase and RNF146.
- Tankyrase inhibits Rab11a activation by suppressing SH3BP5 and SH3BP5L, thereby blocking cyst lumen formation.
- RNF146 promotes lumen formation by reducing Tankyrase levels, which in turn reactivates Rab11a via SH3BP5/SH3BP5L.
Conclusions:
- Epithelial polarity and cyst lumen formation are regulated by a pathway involving Tankyrase, RNF146, SH3BP5, SH3BP5L, and Rab11a.
- Tankyrase acts as an inhibitor of lumenogenesis by targeting SH3BP5 paralogues.
- RNF146 functions as a positive regulator by antagonizing Tankyrase, thereby enabling Rab11a-mediated lumen formation.
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