| 2002 |
Rapostlin was identified as a novel effector protein of Rnd2 GTPase. In vitro binding assays demonstrated that Rapostlin specifically binds Rnd2 (but not other Rho family GTPases) in a GTP-dependent manner. The Rnd2-binding domain of Rapostlin is located between its FCH and SH3 domains. Rapostlin also directly binds microtubules via its amino-terminal FCH domain-containing region. In PC12 cells, Rapostlin induces neurite branching in response to Rnd2, requiring at least the amino-terminal region of Rapostlin. |
Yeast two-hybrid screen, in vitro binding assay, co-expression in PC12 cells with morphological readout |
The Journal of biological chemistry |
High |
12244061
|
| 2004 |
Rapostlin splicing variants (RapostlinL, RapostlinM, RapostlinS) all bind Rnd2 in a GTP-dependent manner. The unique insert region distinguishes their neurite-branching activity: RapostlinM and RapostlinS induce less branching than RapostlinL when co-expressed with Rnd2 in PC12 cells. All variants bind N-WASP in vitro and in vivo via their SH3 domain, and this SH3 domain is essential for branching activity. Rnd2 reduces the RapostlinL–N-WASP interaction but has little effect on RapostlinM or RapostlinS interaction with N-WASP. |
Dot-blot GTP-binding assay, co-expression in PC12 cells, immunoprecipitation, in vitro binding assay |
The Journal of biological chemistry |
Medium |
14732713
|
| 2002 |
Vps4-A (an AAA ATPase family member and central regulator of early endosome trafficking) was identified as a binding partner of Rnd2 by yeast two-hybrid screening, confirmed by in vitro binding and co-immunoprecipitation. Vps4-A associates with both GTP- and GDP-bound forms of Rnd2. When co-expressed with an ATPase-defective Vps4-A mutant (E228Q) that accumulates in early endosomes, Rnd2 is recruited to those early endosomes, suggesting Rnd2 is involved in endosomal trafficking via direct binding to Vps4-A. |
Yeast two-hybrid, in vitro binding assay, co-immunoprecipitation, co-expression/co-localization in HeLa cells |
The Biochemical journal |
Medium |
11931639
|
| 2006 |
Pragmin was identified as a novel effector of Rnd2 GTPase. In vitro and in vivo binding assays showed Pragmin specifically binds Rnd2 (but not other Rho family GTPases) in a GTP-dependent manner. Rnd2-bound Pragmin significantly stimulates RhoA activity and induces cell contraction through RhoA and the Rho-kinase pathway in HeLa cells. In PC12 cells, Pragmin expression inhibits NGF-induced neurite outgrowth in a Rnd2-dependent manner, and Pragmin knockdown by siRNA enhances neurite elongation. Thus Rnd2 can activate RhoA signaling through Pragmin, in contrast to Rnd1 and Rnd3 which inhibit RhoA. |
Yeast two-hybrid, in vitro and in vivo binding assays, RhoA activity assay, siRNA knockdown in PC12 cells, morphological readout |
The Journal of biological chemistry |
High |
16481321
|
| 2003 |
MgcRacGAP was identified as a binding partner of Rnd2 in male germ cells. GST pull-down and co-immunoprecipitation experiments demonstrated a stable Rnd2–MgcRacGAP complex. Both proteins are co-expressed in spermatocytes and spermatids (where classical Rho GTPases RhoA, Rac1, Cdc42 are absent), and they co-localize in the Golgi-derived pro-acrosomal vesicle, suggesting Rnd2 is a physiological partner of MgcRacGAP in male germ cells. |
GST pull-down, co-immunoprecipitation, co-localization by immunofluorescence in germ cells |
The Biochemical journal |
Medium |
12590651
|
| 2005 |
In utero electroporation experiments in mouse embryonic cerebral cortex showed that exogenous expression of wild-type or constitutively active Rnd2, but not a dominant-negative mutant, disturbed morphology and radial migration of pyramidal neurons to upper cortical layers. Rnd2 was expressed by radially migrating cells in the subventricular zone, establishing an in vivo function of Rnd2 activity in pyramidal neuron migration and morphological changes. |
In utero electroporation (wild-type, constitutively active, and dominant-negative Rnd2 constructs), in vivo morphological analysis |
Neuroscience research |
Medium |
16303198
|
| 2008 |
Neurogenin 2 (Neurog2) directly induces Rnd2 expression in newly generated mouse cortical neurons prior to migration. Rnd2 silencing phenocopies the radial migration defect seen in Neurog2-null neurons, and restoring Rnd2 expression in Neurog2-mutant neurons rescues their migratory ability. This places Rnd2 as a direct transcriptional target and key effector of Neurog2 in promoting cortical neuron migration. |
In utero electroporation (shRNA silencing of Rnd2), Neurog2 knockout mouse analysis, rescue experiments by Rnd2 re-expression, transcriptional target validation |
Nature |
High |
18690213
|
| 2011 |
COUP-TFI directly represses Rnd2 expression at the post-mitotic level along the rostrocaudal axis of the neocortex. Loss of COUP-TFI leads to increased Rnd2 expression, delayed radial migration, and morphological defects in callosal projection neurons. Restoring correct Rnd2 levels in COUP-TFI-null brains cell-autonomously rescues radial migration, morphological transitions, axonal elongation, and dendritic arborization defects. |
COUP-TFI knockout mouse analysis, in utero electroporation, Rnd2 rescue experiments in vivo, transcriptional repression assays |
Development (Cambridge, England) |
High |
21965613
|
| 2013 |
The zinc finger transcription factor RP58 (ZNF238) directly represses Rnd2 transcription by binding to a 3'-regulatory enhancer in a sequence-specific fashion. Loss of RP58 in embryonic cortex elevates Rnd2 mRNA and impairs neuronal migration and positioning. Reporter assays showed RP58 repression of Rnd2 is competed by proneural bHLH transcriptional activators. In vivo rescue experiments confirmed that RP58-mediated negative regulation of Rnd2 is critical for cortical cell migration. |
RP58 knockdown in embryonic cortex, reporter assays, chromatin binding/enhancer analysis, in vivo rescue experiments |
Cerebral cortex (New York, N.Y. : 1991) |
High |
24084125
|
| 2015 |
Bacurd2 (BTB-domain containing adaptor for Cul3-mediated RhoA degradation 2) was identified as a novel interacting partner of Rnd2 that binds at Rnd2's C-terminus, an interaction required for its cell migration function. Forced expression or knockdown of Bacurd2 both impair radial neuronal migration and alter immature neuron morphology. Bacurd2 influences the multipolar-to-bipolar transition of radially migrating neurons cell-autonomously. A Bacurd2–Rnd2 chimeric construct experiment suggests the two proteins interact to cooperatively promote radial migration. |
Binding domain mapping, in utero electroporation (overexpression and knockdown), in vivo neuronal morphology analysis, chimeric construct epistasis |
Neural development |
Medium |
25888806
|
| 2020 |
RND2 physically interacts with p38 MAPK and decreases p38 phosphorylation, thereby functioning as an endogenous repressor of the p38 MAPK phosphorylation complex in glioblastoma cells. Forced RND2 expression represses p38 MAPK signaling, inhibiting autophagy and apoptosis. Conversely, RND2 downregulation enhances p38 signaling and promotes autophagy and apoptosis. Inhibition of p38 phosphorylation abolishes the pro-apoptotic/autophagic effects of RND2 deficiency. |
Co-immunoprecipitation (RND2–p38 interaction), western blot (p38 phosphorylation), flow cytometry/TUNEL/LC3B assays (apoptosis/autophagy), overexpression and knockdown in GBM cells, intracranial xenograft in vivo |
Journal of experimental & clinical cancer research : CR |
Medium |
32867814
|
| 2021 |
Rnd2 is required for survival, positioning, somatodendritic morphogenesis, and functional maturation of adult-born dentate granule neurons, as shown by retrovirus-based loss-of-function in vivo. These functions are largely specific to adult neurogenesis; deletion in neonatally-born granule neurons only affects dendritogenesis. Suppression of Rnd2 in adult-born neurons increases anxiety-like behavior. |
Retrovirus-mediated Rnd2 loss-of-function in vivo (adult and neonatal hippocampal neurogenesis), morphological analysis, behavioral assays |
Molecular psychiatry |
Medium |
34561615
|
| 2021 |
Rnd2 plays biphasic roles in oligodendrocyte myelination: oligodendrocyte-specific Rnd2 knockout mice show decreased myelin thickness at the onset of myelination but increased myelin thickness later. Correspondingly, phosphorylation of Rho kinase and its substrate Mbs (a signaling unit negatively regulating myelination) is higher at early myelination onset and lower later in Rnd2 KO mice. Oligodendrocyte-specific Rnd2 transgenic mice confirm the biphasic role. Thus Rnd2 positively regulates early myelination and negatively regulates later myelination through the Rho kinase/Mbs pathway. |
Oligodendrocyte-specific Rnd2 knockout and transgenic mice, myelin thickness measurements, western blot for Rho kinase and Mbs phosphorylation |
Molecular biology of the cell |
High |
33596091
|
| 2023 |
Knockdown of Rnd2 (via CRISPR/CasRx or RNAi) in oligodendroglial FBD-102b cells slows morphological differentiation. Downstream signaling involves Prag1 (Pragmin) and Fyn kinase; knockdown of Prag1 or Fyn also slows differentiation, and Rnd2 or Prag1 knockdown decreases Fyn phosphorylation (critical for its activation). This establishes a Rnd2→Prag1→Fyn kinase signaling axis controlling oligodendroglial morphological differentiation. |
CRISPR/CasRx and RNAi knockdown, western blot for Fyn phosphorylation, morphological differentiation assay in FBD-102b cell line |
Neurology international |
Medium |
38251051
|
| 1999 |
RhoN (RND2) was cloned from rat spinal cord and identified as a novel Rho subfamily small GTPase specifically expressed in neurons and hepatic stellate cells. Unlike classical Rho proteins, RhoN is not susceptible to ADP-ribosylation by C3 botulinum toxin, indicating insensitivity to C3 toxin-mediated inactivation. |
cDNA cloning, RNA blot hybridization, in situ hybridization, ADP-ribosylation assay with C3 botulinum toxin, genomic mapping |
Brain research. Molecular brain research |
Medium |
10101234
|