Chapter Twenty-Four - MICAL-Like1 in Endosomal Signaling

Abstract

Small GTPase Rabs are required for membrane protein sorting/delivery to precise membrane domains. Rab13 regulates tight junction assembly and polarized membrane transport in epithelial cells. Using yeast two-hybrid screen, we identified MICAL-like1 (MICAL-L1), a protein that interacts with GTP-bound Rab13 and shares a similar domain organization with MICAL protein family. MICAL-L1 has a calponin homology, Lin11, Isl-1 & Mec-3 (LIM), proline-rich, and coiled-coil domains. It is associated with late and recycling endosomes. Time-lapse video microscopy shows that GFP-Rab7 and cherry-MICAL-L1 are present within vesicles that move rapidly in the cytoplasm. Depletion of MICAL-L1 by short hairpin RNA does not alter the distribution of tight junction proteins, but affects the trafficking of epidermal growth factor receptor (EGFR). Overexpression of MICAL-L1 leads to the accumulation of EGFR in late endosomal compartments. In contrast, knocking down MICAL-L1 results in the distribution of internalized EGFR in vesicles spread throughout the cytoplasm and promotes its degradation. Our data show that MICAL-L1 inhibits EGFR degradation, suggesting that MICAL-L1 is involved in sorting/targeting the receptor to the recycling pathway. They provide novel insights into MICAL-L1/Rab protein complex that can regulate EGFR trafficking/signaling.