| 2009 |
MICAL-L1 directly interacts with EHD1 and Rab8a, colocalizing on tubular recycling endosomes. The MICAL-L1 C-terminal coiled-coil region is necessary and sufficient for localization to tubular membranes. MICAL-L1 depletion causes loss of EHD1-Rab8a interaction and absence of both proteins from tubular membranes, implicating MICAL-L1 as a scaffold/Rab effector that recruits and links EHD1 and Rab8a to tubular recycling endosomes to facilitate receptor recycling. |
Co-immunoprecipitation, live-cell imaging, siRNA knockdown, domain truncation experiments (coiled-coil region), transferrin and integrin receptor recycling assays |
Molecular biology of the cell |
High |
19864458
|
| 2011 |
MICAL-L1 interacts with Rab35 and Arf6 in vivo. Active Rab35 overexpression impairs MICAL-L1 recruitment to tubular recycling endosomes, while Rab35 depletion enhances MICAL-L1 localization there. Arf6 forms a complex with MICAL-L1 and contributes to its endosomal recruitment. This positions Rab35 as a critical upstream regulator of MICAL-L1 and Arf6, with MICAL-L1 and Arf6 acting upstream of Rab8a. |
Co-immunoprecipitation, siRNA knockdown, overexpression of GTPase mutants (active/inactive), fluorescence microscopy |
Traffic (Copenhagen, Denmark) |
High |
21951725
|
| 2013 |
MICAL-L1 and the BAR-domain protein syndapin2 both bind phosphatidic acid (a novel lipid component of recycling endosomes). Direct protein-protein interaction between MICAL-L1 and syndapin2 stabilizes membrane association. Phosphatidic acid in liposomes enhances syndapin2-mediated membrane tubulation in vitro, supporting a model in which MICAL-L1 and syndapin2 cooperate to nucleate tubular recycling endosome biogenesis. |
Lipid-binding assays (phosphatidic acid), in vitro liposome tubulation assay with recombinant syndapin2, co-immunoprecipitation, siRNA knockdown |
Molecular biology of the cell |
High |
23596323
|
| 2014 |
During NGF-induced neurite outgrowth, Rab35 recruits MICAL-L1 to Arf6-positive recycling endosomes, and MICAL-L1 in turn scaffolds Rab8, Rab13, and Rab36 to those endosomes. Knockdown of Rab36 impaired recruitment of the Rab36 effector JIP4 to recycling endosomes and inhibited neurite outgrowth without affecting Rab8 or Rab13 localization, demonstrating non-redundant downstream Rab functions. |
siRNA knockdown of Rab35, MICAL-L1, Rab8, Rab13, Rab36; fluorescence microscopy; neurite outgrowth assays in NGF-treated PC12 cells |
Biology open |
Medium |
25086062
|
| 2014 |
GRAF1 forms a complex with MICAL-L1 and EHD1 at tubular recycling endosomes. GRAF1 overexpression causes vesiculation of MICAL-L1-positive tubules; GRAF1 depletion impairs tubular recycling endosome vesiculation and delays receptor recycling. Co-addition of purified EHD1 and GRAF1 in a semi-permeabilized cell assay produces synergistic tubule vesiculation. |
Co-immunoprecipitation, siRNA knockdown, overexpression, semi-permeabilized cell vesiculation assay with purified proteins |
Frontiers in cell and developmental biology |
Medium |
25364729
|
| 2014 |
MICAL-L1 depletion results in increased binucleated cells due to impaired recycling endosome transport during late cytokinesis, and also causes aberrant chromosome alignment and lagging chromosomes independently of EHD1, indicating an EHD1-independent role for MICAL-L1 earlier in mitosis. Both MICAL-L1 and EHD1 differentially influence microtubule dynamics during early and late mitosis. |
siRNA knockdown of MICAL-L1 and EHD1, binucleation assays, chromosome alignment imaging, microtubule dynamics analysis |
Traffic (Copenhagen, Denmark) |
Medium |
25287187
|
| 2014 |
MICAL-L1 is required for growth-factor- and integrin-induced Src activation and transport to the cell periphery. MICAL-L1 depletion impairs focal adhesion turnover, cell spreading, and migration. MICAL-L1-mediated recruitment of EHD1 to Src-containing recycling endosomes is required for Src release from the perinuclear recycling compartment in response to growth factor stimulation. |
siRNA knockdown of MICAL-L1 and EHD1, co-localization by fluorescence microscopy, Src activation assays, focal adhesion turnover assays, migration assays in HeLa cells and human fibroblasts |
Journal of cell science |
Medium |
24481818
|
| 2017 |
MICALL1 is a transcriptional target of p53; functional p53-binding motifs were identified ~3000 bp upstream of the MICALL1 gene. In response to DNA damage, MICALL1 co-localizes with RAB8A and CD2AP at tubular recycling endosomes; siRNA depletion of p53 or MICALL1 prevents this localization, demonstrating that p53 regulates tubular recycling endosome biogenesis via MICALL1. |
Multi-omics (mass spectrometry, cDNA microarray), p53 binding motif identification, siRNA knockdown, co-localization by fluorescence microscopy |
International journal of oncology |
Medium |
28714518
|
| 2019 |
MICAL-L1 localizes to cilia and centrosomes (including non-ciliated cells). MICAL-L1 knockdown impairs ciliogenesis similarly to EHD1 knockdown, prevents CP110 removal from the mother centriole, and causes EHD1 to fail to localize to basal bodies. Mass spectrometry and direct binding assays identified α-tubulin–β-tubulin heterodimers and γ-tubulin as direct MICAL-L1 interaction partners, supporting a model in which centriolar tubulins anchor MICAL-L1 to recruit EHD1 for ciliogenesis. |
siRNA knockdown, fluorescence microscopy (localization to cilia/centrosomes), mass spectrometry, direct protein-binding assays (tubulin interactions) |
Journal of cell science |
Medium |
31615969
|
| 2021 |
PACSIN2 recruits EHD4 and MICAL-L1 to the rear end of asymmetric adherens junctions in endothelial cells, forming a recycling endosome-like tubular structure. This junctional PACSIN2/EHD4/MICAL-L1 complex controls local VE-cadherin trafficking and coordinates polarized endothelial migration and angiogenesis. |
Co-immunoprecipitation, fluorescence microscopy, siRNA/shRNA knockdown, VE-cadherin trafficking assays, angiogenesis sprouting assays |
Nature communications |
Medium |
33972531
|
| 2021 |
MICAL-L1 depletion impairs delivery of a subset of cargo proteins to the cell surface. The MICAL-L1 RBD domain contributes to PACSIN-mediated membrane tubulation in vitro. Two hydrophobic residues at the MICAL-L1 C-terminus are important for phosphatidic acid binding and for association with membrane tubules. MICAL-L1 associates with Golgi apparatus markers and recycling endosomes. |
shRNA knockdown, synchronized secretion assay, in vitro membrane tubulation assay with recombinant MICAL-L1-RBD, confocal and STORM super-resolution microscopy, site-directed mutagenesis of C-terminal hydrophobic residues |
Biology open |
Medium |
34100897
|
| 2024 |
MICAL-L1 directly interacts with FCHSD2 and recruits it to the endosomal membrane. FCHSD2 recruitment by MICAL-L1 is required for ARP2/3-mediated branched actin generation, endosome fission, and receptor recycling. Because MICAL-L1 recruits FCHSD2 prior to EHD1, MICAL-L1 orchestrates the sequential steps of endosomal fission by bridging early actin-driven constriction (via FCHSD2/ARP2/3) and subsequent nucleotide hydrolysis/fission (via EHD1). |
Co-immunoprecipitation, siRNA knockdown of MICAL-L1 and FCHSD2, ARP2/3 actin branching assays, endosome fission assays, receptor recycling assays |
Molecular biology of the cell |
Medium |
39382837
|
| 2025 |
CD2AP and CIN85 are novel constituents of tubular recycling endosomes recruited by MICAL-L1 via their SH3 domains. Depletion of either CD2AP or CIN85 impairs recycling endosome function. MICAL-L1 scaffolding of these proteins likely impacts recycling through effects on the actin cytoskeleton. |
Co-immunoprecipitation (SH3 domain interactions), siRNA knockdown of CD2AP and CIN85, receptor recycling assays, fluorescence microscopy |
Traffic (Copenhagen, Denmark) |
Medium |
40740057
|
| 2014 |
MICAL-L1 is necessary for ETEN (elongated tubular endosomal network) remodeling originating from the endocytic recycling compartment in human dendritic cells, as demonstrated by knockdown experiments. |
siRNA knockdown, fluorescence microscopy |
Communicative & integrative biology |
Low |
26478765
|