| 2011 |
MICAL3 interacts with both Rab8A and ELKS, linking these two proteins. Rab8A associates with exocytotic vesicles in a Rab6-dependent manner, and MICAL3 acts as a scaffold connecting Rab8A and ELKS at the cell cortex. Expression of a MICAL3 mutant with an inactive monooxygenase domain caused strong accumulation of secretory vesicles docked at the cell cortex that failed to fuse with the plasma membrane, indicating that MICAL3's monooxygenase activity is required for vesicle-docking complex remodeling and fusion. |
Co-immunoprecipitation, live-cell imaging of vesicle dynamics, dominant-negative monooxygenase mutant expression, FRAP |
Current biology : CB |
High |
21596566
|
| 2005 |
MICAL3 interacts with Rab1 in yeast two-hybrid and GST pulldown experiments. MICAL3 localizes to a microtubule-associated compartment, as treatment with the microtubule-depolymerizing drug nocodazole disrupts this localization, indicating a link between MICAL3 and the microtubule cytoskeleton. |
Yeast two-hybrid, GST pulldown, immunofluorescence with nocodazole treatment |
Biochemical and biophysical research communications |
Medium |
15694364
|
| 2007 |
Knockdown of MICAL3 in chick motor neurons via RNAi caused ectopic positioning of motor neuron cell bodies in the peripheral nervous system, placing MICAL3 downstream of Neuropilin-2/Plexin-A2 semaphorin signaling as a cytoplasmic effector that controls somal positioning through cytoskeletal reorganization. |
RNA interference in chick embryo, analysis of motor neuron positioning |
Neural development |
Medium |
17971221
|
| 2012 |
MICAL3 (as MICAL-3) was shown to interact with Rab1 and Rab35 effector binding is only slightly inhibited by phosphocholination, in contrast to GEF and GDI interactions which are strongly inhibited, demonstrating that MICAL-3 functions as a Rab effector protein. |
Biochemical binding assays with phosphocholinated Rab proteins, quantitative interaction measurements |
The EMBO journal |
Medium |
22307087
|
| 2012 |
Human MICAL3, like MICAL1 and MICAL2, is required for normal actin stress fiber organization in non-neural cells, and this actin-regulatory function depends on its generation of reactive oxygen species (ROS) via the monooxygenase domain. |
siRNA knockdown, ROS inhibitor treatment, immunofluorescence of actin stress fibers |
Journal of cell science |
Medium |
22331357
|
| 2015 |
MICAL3 was identified as part of a NINL-associated interactome at the base of cilia, and interacts with Rab8 to regulate vesicle docking and fusion for cilia-directed cargo delivery. Genetic interaction between NINL and CC2D2A was demonstrated in zebrafish, and MICAL3 was linked to this pathway. |
Co-immunoprecipitation/interactome analysis, zebrafish morpholino knockdown, immunolocalization |
PLoS genetics |
Medium |
26485645
|
| 2016 |
MICAL3 forms a direct complex with the centralspindlin component MKLP1, is recruited to the central spindle and midbody during cytokinesis, and targets ELKS and Rab8A-positive vesicles to the midbody. Knockout of MICAL3 increases cytokinetic failure frequency and delays abscission. This scaffolding function is independent of MICAL3's enzymatic monooxygenase activity. |
Cross-linking mass spectrometry, MICAL3 knockout, cell biological assays (cytokinesis failure quantification, abscission timing), immunofluorescence |
The Journal of biological chemistry |
High |
27528609
|
| 2017 |
Cross-linking mass spectrometry combined with deletion analysis defined the minimal binding domains of the MICAL3/ELKS/Rab8A complex involved in exocytosis, providing structural constraints on this protein assembly. |
Cross-linking mass spectrometry (XL-MS), deletion analysis, biochemical interaction assays |
Scientific reports |
Medium |
29044157
|
| 2018 |
Human MICAL3 directly associates with F-actin, which activates its catalytic activity. MICAL3 uses NADPH-dependent redox activity to oxidize actin methionine residues M44/M47, dismantling filaments and limiting polymerization. MsrB/SelR reductase enzymes counteract MICAL3's effect on F-actin both in vitro and in vivo. |
In vitro F-actin binding and disassembly assays, NADPH consumption assays, genetic experiments in Drosophila, mass spectrometry of oxidized actin residues |
Scientific reports |
High |
29343822
|
| 2018 |
Semaphorin 3 stimulation induces interaction among MICAL3, CRMP2, and Numb in breast cancer stem-like cells. MICAL3 monooxygenase activity (MO activity) is required for this interaction, for Numb protein accumulation, and for symmetric cell division. MICAL3 knockdown decreases sphere formation and shifts CSCs from symmetric to asymmetric division. |
Co-immunoprecipitation, MICAL3 siRNA knockdown, sphere formation assay, division mode quantification (symmetric vs. asymmetric) |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
30587593
|
| 2020 |
Crystal structure of human MICAL3 FMO and calponin-homology (CH) domains was solved. MICAL3 contains an FAD/NADP-binding Rossmann-fold domain for monooxygenase activity. Kinetic analysis showed that F-actin dramatically increases MICAL3 catalytic efficiency only when the CH domain is present. Mutation of Glu213 in the FMO domain and Arg530 in the CH domain abolished this F-actin-stimulated catalytic activation. |
X-ray crystallography, kinetic enzyme assays, site-directed mutagenesis (E213A, R530A) |
IUCrJ |
High |
31949908
|
| 2024 |
CHK1 physically interacts with MICAL3 in mouse zygotes, as identified by co-immunoprecipitation and mass spectrometry. Gain-of-function CHK1 mutants enhance interaction with MICAL3 and increase MICAL3 enzymatic activity, causing excessive F-actin depolymerization that disrupts pronuclear envelope breakdown. |
Co-immunoprecipitation and mass spectrometry of ~6000 mouse zygotes, gain-of-function CHK1 mutant analysis, F-actin imaging |
EMBO reports |
Medium |
39358552
|