| 2002 |
Rab34 is associated primarily with the Golgi apparatus, and expression of wild-type or GTP-restricted (but not GDP-restricted) Rab34 causes spatial redistribution of lysosomes from the periphery to the peri-Golgi region. This activity requires membrane association via prenylation and direct interaction with RILP (Rab-interacting lysosomal protein), dependent on Lys82 in the switch I region. |
Overexpression of wild-type and mutant Rab34 constructs, GST pull-down, direct binding assays, site-directed mutagenesis (K82), immunofluorescence microscopy |
Molecular biology of the cell |
High |
12475955
|
| 2002 |
Rab34/Rah localizes to membrane ruffles and nascent macropinosomes (co-localizing with actin), has extremely low intrinsic GTPase activity in vitro but appreciable activity in vivo (suggesting a GAP), and promotes macropinosome formation downstream of Rac1 and WAVE2. |
Cloning and in vitro GTPase assay, fluorescence microscopy, dominant-negative and constitutively active mutant overexpression, PDGF/phorbol ester stimulation |
The Journal of biological chemistry |
High |
12446704
|
| 2003 |
RILP interacts selectively with GTP-bound (wild-type and GTP-restricted) but not GDP-restricted Rab34, as demonstrated by yeast two-hybrid and GST pull-down. A 62-residue region (aa272-333) unique to RILP is necessary for regulating lysosomal morphology and for interaction with both Rab7 and Rab34. |
Yeast two-hybrid, GST pull-down, chimeric protein domain-transfer experiments, overexpression and morphological analysis |
Molecular biology of the cell |
High |
14668488
|
| 2005 |
RILP, the shared effector of Rab7 and Rab34, is capable of self-interaction (homodimerization), as demonstrated by yeast two-hybrid and co-immunoprecipitation in HeLa cells. |
Yeast two-hybrid, co-immunoprecipitation in HeLa cells |
Biochemical and biophysical research communications |
Medium |
15996637
|
| 2005 |
Hmunc13 (a diacylglycerol-binding protein) is an effector of GTP-bound Rab34 at the Golgi; interaction requires the MHD-2 domain of hmunc13 and is dependent on the GTP-bound state of Rab34 (Q111L mutant), identified by bacterial two-hybrid screen and confirmed by co-immunoprecipitation. |
Bacterial two-hybrid screen, co-immunoprecipitation, GST pull-down with GTP/GDP-loaded mutants, radioactive GTP overlay assay |
Traffic |
Medium |
16138900
|
| 2005 |
GTP overlay assay confirms that wild-type and GTP-restricted Rab34 bind GTP in vitro; K82 in Rab34 is a key residue required for RILP interaction and for lysosome redistribution in cells. |
GTP overlay assay, GST pull-down, site-directed mutagenesis, mammalian cell overexpression |
Methods in enzymology |
High |
16473629
|
| 2007 |
CVB (coxsackievirus B) entry across epithelial tight junctions requires Rab34 (and Rab5) activity; both occludin internalization into macropinosomes and CVB infection are blocked by dominant-negative Rab34, indicating Rab34 functions in macropinocytic viral entry. |
Dominant-negative Rab34 expression, siRNA depletion, endocytosis/infection assays in polarized epithelial cells, inhibitors of macropinocytosis |
Cell host & microbe |
High |
18005733
|
| 2007 |
Rab34 localizes to the Golgi stack and functions in intra-Golgi transport (downstream of ER, upstream of trans-Golgi network exit); depletion by dominant-negative Rab34 or RNAi blocks VSVG-GFP transport from Golgi to plasma membrane without affecting ER-to-medial Golgi traffic, as shown by endoglycosidase H resistance assay and brefeldin A treatment. |
Immunoelectron microscopy, immunocytochemistry, RNAi knockdown, dominant-negative overexpression, VSVG-GFP trafficking assay, EndoH resistance assay, brefeldin A treatment |
Molecular biology of the cell |
High |
17881736
|
| 2009 |
Rab34 and munc13-2 form a functional complex at the Golgi that mediates protein secretion; siRNA knockdown of either munc13-2 or Rab34 abolishes high glucose-induced VSVG-GFP secretion, and munc13-2 with deleted MHD2 cannot rescue, establishing a Rab34–munc13-2 axis in regulated secretion. |
siRNA knockdown, VSVG-GFP secretion assay, MHD2 deletion mutant transfection, fibronectin secretion measurement |
American journal of physiology. Cell physiology |
Medium |
19641095
|
| 2012 |
Rab34 mediates phagolysosome biogenesis through recruitment of Munc13-2; Rab34 knockdown impairs lysosome-phagosome fusion independently of Rab7, and Rab34-mediated phagosome maturation is critical for mycobacterial killing. |
siRNA knockdown, overexpression of active Rab34, phagolysosome fusion assay, mycobacterial survival assay, co-localization studies |
Proceedings of the National Academy of Sciences of the United States of America |
High |
23197834
|
| 2016 |
Folliculin (FLCN) promotes peri-nuclear lysosome clustering via its C-terminal DENN domain interacting directly with the Rab34 effector RILP; using purified recombinant proteins, FLCN-DENN does not act as a GEF for Rab34 but loads active Rab34 onto RILP to form a ternary complex, restricting lysosome motility under starvation. |
Purified recombinant protein binding assay, GEF assay, knockdown/overexpression, live-cell imaging, lysosome motility analysis |
EMBO reports |
High |
27113757
|
| 2018 |
Rab34 localizes to cilia in vivo; Rab34 mutation in mice impairs preciliary vesicle fusion to form ciliary vesicles and blocks mother centriole migration to the plasma membrane, resulting in reduced ciliogenesis, polydactyly, cleft lip/palate, and failure of Hedgehog signaling (reduced Gli3 processing). |
Mouse knockout/mutation, immunofluorescence, electron microscopy, Gli3 processing assay, phenotypic analysis |
Journal of cell science |
High |
30301781
|
| 2018 |
Rab34 binds to the cytoplasmic tail of integrin β3 and prevents its degradation; EGF induces translocation of Rab34 to membrane ruffles, enhanced by Src kinase; Src phosphorylates Rab34 at Y247, and a phosphomimetic mutant (Y247D) promotes cell migration, invasion, integrin β3 endocytosis, and recycling. |
Co-immunoprecipitation, shRNA knockdown, overexpression of phosphomimetic/phospho-dead mutants, EGF stimulation, in vitro kinase assay, cell migration/invasion assays |
Oncogene |
High |
29622794
|
| 2020 |
A comprehensive siRNA screen identified Rab34 as essential for serum starvation-induced ciliogenesis in hTERT-RPE1, NIH/3T3, and MCF10A cells (but not MDCK-II cysts); a unique long N-terminal region (aa1-49) of Rab34, rather than the switch II region, is required for this function. |
Genome-wide siRNA knockdown screen (62 Rabs), Rab34 KO by CRISPR, deletion/mutation analysis, ciliogenesis assay |
The Journal of biological chemistry |
High |
32669361
|
| 2021 |
Rab34 is specifically required for the intracellular (but not extracellular/surface) ciliogenesis pathway; it marks the ciliary sheath, a unique sub-domain of assembling intracellular cilia, and is required for ciliary vesicle formation at the mother centriole. GTP binding and turnover by Rab34 are both required for ciliogenesis, modulated by divergent residues in its GTPase domain. |
Rab34 KO in multiple cell lines, live-cell imaging, electron microscopy, GTPase activity assays, domain/residue mutagenesis, MDCK extracellular pathway comparison |
Current biology : CB |
High |
33989527
|
| 2021 |
Proximity biotinylation with Ift27 as bait identified Rab34 as a ciliary protein; Rab34 localizes near the mother centriole and is required for ciliary vesicle formation at an early step. In fibroblasts (using internal ciliogenesis pathway), Rab34 loss blocks ciliogenesis; in epithelial IMCD3 cells at low density (using internal pathway), Rab34 is also required. |
Proximity biotinylation (BioID), Rab34 KO/knockdown, immunofluorescence, ciliogenesis assay across cell types and densities |
Current biology : CB |
High |
33989524
|
| 2021 |
The N-terminal LPQ sequence (amino acids 16-18) of Rab34 is specifically required for ciliogenesis in hTERT-RPE1 cells; a Rab34 mutant with LPQ→AAA substitution fails to rescue the Rab34-KO ciliogenesis defect. |
Rab34 KO rescue assay, deletion analysis, site-directed mutagenesis (LPQ→AAA), ciliogenesis assay |
Small GTPases |
Medium |
33860735
|
| 2023 |
Pathogenic bi-allelic missense variants in RAB34 clustered near the C-terminus cause oral-facial-digital syndrome (OFDS-RAB34) with loss of ciliogenesis function; cells expressing mutant RAB34 show significant defects in cilium assembly, with some variants retaining mother centriole recruitment but failing subsequent steps of intracellular ciliogenesis. |
Exome sequencing, patient-derived cell functional assays, ciliogenesis assay, immunofluorescence |
Human molecular genetics |
Medium |
37384395
|
| 2024 |
DENND6A is a GEF for Rab34; activated by Arl8b on peripheral lysosomes, DENND6A activates Rab34, which then recruits a RILP/dynein complex to lysosomes to promote retrograde transport. Loss of DENND6A impairs autophagic flux, placing Arl8b→DENND6A→Rab34→RILP/dynein as a regulatory axis controlling nutrient-dependent juxtanuclear lysosome repositioning. |
Cell-based GEF screen, GEF activity assay, co-immunoprecipitation, pulldown with purified proteins, lysosome positioning assay, autophagic flux assay, epistasis experiments |
Nature communications |
High |
38296963
|
| 2024 |
Rab34 regulates type I collagen trafficking from the ER to the Golgi and is required for Golgi cisternae integrity; in adipocytes, Rab34 translocates from the Golgi to lipid droplets during lipid droplet biogenesis, where it controls lipolysis through interaction with the E1-ubiquitin ligase UBA1, which ubiquitinates FABP5 for proteasomal degradation. At the Golgi, Rab34 regulates adiponectin trafficking and oligomerization. |
Immunofluorescence with organelle markers, siRNA knockdown, overexpression, proteomic interactome analysis, adiponectin secretion/oligomerization assay, lipolysis assay, co-immunoprecipitation |
Journal of biomedical science |
Medium |
38183057
|
| 2024 |
Rab34 is required for cilia formation and cilia-mediated Hedgehog signaling in craniofacial development; it also has a non-ciliary function in regulating type I collagen trafficking from the ER to the Golgi, impacting osteogenesis. |
Mouse conditional knockout, immunofluorescence, Hedgehog signaling assays, collagen trafficking assay |
Biochemical and biophysical research communications |
Medium |
38852507
|
| 2017 |
Salmonella effector SopD2 binds Rab34 and modulates its function; depletion of Rab34 delays maturation of the Salmonella-containing vacuole (SCV) and inhibits intracellular S. typhimurium growth, establishing Rab34 as a host factor required for SCV maturation. |
Co-immunoprecipitation, siRNA knockdown, intracellular bacterial growth assay, SCV maturation assay |
Cell biology international |
Medium |
28185347
|
| 2023 |
PSMB1 (proteasome subunit beta type-1) binds directly to RAB34 and promotes its proteasome-dependent degradation, leading to inactivation of the MEK/ERK signaling pathway; Kinetin enhances the PSMB1-RAB34 interaction and accelerates RAB34 degradation. |
Co-immunoprecipitation, proteasome inhibitor assay, MEK/ERK phosphorylation western blot, PDX and liver metastasis xenograft models, computer-aided drug design |
Cancer letters |
Medium |
38159835
|
| 2022 |
Rab34 acts as a negative regulator of osteoclast differentiation by promoting lysosomal proteolysis of osteoclastogenic surface receptors c-fms and RANK via the early endosome–late endosome–lysosome axis, thereby attenuating c-fos/NFATc1 transcriptional activity; Rab34 also modulates secretion of lysosomal proteases (MMP9, Cathepsin K) at ruffled borders. |
siRNA knockdown, overexpression, osteoclast differentiation assay (RAW-D cells and bone marrow-derived macrophages), receptor degradation assay, transcription factor activity assay |
Cell biochemistry and function |
Medium |
35285960
|