| 1988 |
YPT1/Rab1A protein is associated with the secretory machinery; yeast ypt1-1 mutation causes accumulation of membranes and vesicles and a partial secretion defect; immunofluorescence showed the mammalian counterpart localizes to the Golgi apparatus, consistent with a role directing intracellular vesicle traffic. |
Genetic analysis (ypt1-1 mutation phenotype), immunofluorescence with affinity-purified antibody in yeast and mouse cells |
Cell |
High |
3127057
|
| 1986 |
YPT1 protein is essential for cell growth; it binds GTP specifically and GTP binding is required for intracellular function; a dominant-lethal N121I substitution (site-directed mutagenesis) caused binucleated cells and abnormal spindles, demonstrating an essential function in cell cycle/microtubule organization. |
GAL10-regulated expression (depletion), site-directed mutagenesis, GTP-binding assay, immunofluorescence of microtubules |
Cell |
High |
3094963
|
| 1989 |
Ypt1p is required for ER-to-Golgi protein transport in vitro; ypt1 mutations specifically abolish Golgi complex function without affecting ER activity; wild-type cytosol can restore mutant Golgi function, placing Ypt1p at an early vesicular transport step. |
In vitro transport assay with ypt1 mutant membranes, complementation with wild-type cytosol, genetic epistasis with other secretion genes |
The Journal of cell biology |
High |
2504726
|
| 1990 |
Ypt1p is required for ER-to-Golgi transport in cell-free extracts; anti-Ypt1 antibodies block transport; recombinant Ypt1p rescues the inhibition; ypt1-1 extract transport is restored by wild-type cytosol. Ypt1p acts independently of Ca2+ at a distinct step. |
In vitro transport assay, antibody inhibition, recombinant protein rescue, Ca2+ buffering experiments |
Proceedings of the National Academy of Sciences of the United States of America |
High |
2104983
|
| 1991 |
Ypt1p mediates the attachment or fusion step of secretory vesicles with the acceptor compartment (cis-Golgi); blocking Ypt1p activity causes accumulation of transport vesicles; Ypt1p was detected on the outer surface of these vesicles. |
Cell-free ER-to-Golgi transport reconstitution, antibody blocking, vesicle intermediate accumulation assay, immunolocalization on vesicles |
Science (New York, N.Y.) |
High |
1904626
|
| 1991 |
Suppressors of YPT1 deletion (SLY1-20, SLY2, SLY12, SLY41) restore ER-to-Golgi transport in Ypt1p-deficient cells; SLY2 and SLY12 encode synaptobrevin-like proteins; SLY1p is essential and acts as a single-copy suppressor, revealing the downstream machinery of the Ypt1p-regulated transport step. |
Multicopy/single-copy suppressor screens, invertase/CPY processing assays, genetic epistasis, DNA sequencing |
Molecular and cellular biology |
High |
1903839 1990290
|
| 1988 |
YPT1 protein requires at least one of its two C-terminal cysteine residues for palmitic acid (palmitoyl) modification and for membrane association; the non-palmitoylated mutant lacking both cysteines is exclusively soluble and non-functional. |
[3H]palmitic acid metabolic labeling, subcellular fractionation, site-directed mutagenesis, gene replacement |
The EMBO journal |
High |
3042385
|
| 1991 |
Ypt1p membrane attachment depends on BET2 (a Rab geranylgeranyltransferase subunit homologous to DPR1/RAM1); bet2 mutants fail to attach Ypt1p and Sec4p to membranes, establishing prenylation as required for Ypt/Rab membrane association. |
Genetic analysis, DNA sequencing of BET2, membrane fractionation of Ypt1p/Sec4p in bet2 mutants |
Nature |
High |
1903184
|
| 1994 |
Rab geranylgeranyltransferase (RabGGTase)/Rab escort protein catalyzes geranylgeranylation of both C-terminal adjacent cysteines of Rab1A (–XXCC motif), as demonstrated by structural analysis of in vitro prenylated protein. |
In vitro prenylation assay with recombinant RabGGTase, [3H]geranylgeranyl pyrophosphate incorporation, tryptic peptide HPLC, electrospray mass spectrometry |
Proceedings of the National Academy of Sciences of the United States of America |
High |
7991565
|
| 1995 |
Ypt1p is essential for two sequential steps of the yeast secretory pathway: ER-to-cis-Golgi and cis-to-medial-Golgi transport, but not for later steps; this was established using a tight temperature-sensitive allele (ypt1-A136D) and glycoprotein processing assays. |
Temperature-sensitive mutant analysis (ypt1-A136D), glycoprotein processing assays (invertase, CPY, Gas1p), kinetic secretion analysis |
The Journal of cell biology |
High |
7593181
|
| 1993 |
Specificity of Ypt1 versus Sec4 function is encoded in a 9-residue segment corresponding to loop L7 (and a 24-residue segment including the effector region, loop L2) of the GTPase; substitution of these segments of Sec4 with Ypt1 sequences converts Sec4 into a protein that can perform Ypt1 functions. |
Chimeric protein construction, in vivo complementation assays in yeast |
Nature |
High |
8464499
|
| 2000 |
The TRAPP complex is a guanine nucleotide exchange factor (GEF) for Ypt1p (and Ypt31/32); GST-tagged TRAPP subunits Bet3p or Bet5p precipitate GEF activity that stimulates GDP release and GTP uptake on Ypt1p; the dominant-negative Ypt1-D124N mutant inhibits TRAPP GEF activity, suggesting in vivo relevance. |
GST pulldown of TRAPP subunits, GDP-release and GTP-uptake assays in vitro, gel-filtration of GEF activity (>670 kDa), dominant-negative inhibition, overexpression studies |
Molecular biology of the cell |
High |
11102533
|
| 2003 |
Crystal structure of monoprenylated Ypt1:RabGDI complex at 1.5 Å resolution reveals the structural basis for GDI-mediated inhibition of nucleotide release; isoprenoid binding requires a conformational change opening a hydrophobic cavity in GDI domain II. |
Chemical synthesis of prenylated peptide, protein engineering, X-ray crystallography at 1.5 Å |
Science (New York, N.Y.) |
High |
14576435
|
| 2006 |
Structure of doubly geranylgeranylated Ypt1:GDI complex shows one geranylgeranyl moiety deeply buried in GDI domain II hydrophobic pocket while the other is more solvent-exposed; biophysical measurements provide thermodynamic model for GDI- and REP-mediated Rab membrane interaction. |
Synthesis of doubly prenylated protein, X-ray crystallography, biophysical measurements (ITC, fluorescence) |
The EMBO journal |
High |
16395334
|
| 1995 |
A Ypt1p guanine nucleotide exchange factor (GEF) activity is required for ER-to-Golgi transport; dominant-negative Ypt1p-D124N blocks transport by sequestering the GEF; XTP-dependent rescue of Ypt1-D124N confirms the nucleotide-free form sequesters GEF, demonstrating an essential role for the exchange factor in vesicular transport. |
In vitro ER-to-Golgi transport assay, dominant-negative and nucleotide-specificity mutant proteins, GEF activity assay (GDP release, GTP uptake) |
The Journal of cell biology |
High |
7657691
|
| 1991 |
The effector domain of Ypt1p (residues 37–45) is required for function; mutations I41M and D44N are loss-of-function; a GTPase-activating protein (yptGAP) partially purified from porcine liver stimulates GTPase activity of Ypt1p specifically (not H-Ras) and interacts with the effector domain; I41M and D44N mutant proteins show reduced or absent GAP responsiveness. |
Site-directed mutagenesis and gene replacement in yeast, CPY/invertase processing assays, partial purification of yptGAP, GTPase stimulation assays |
The EMBO journal |
High |
2009858
|
| 1998 |
GTP hydrolysis is not essential for Ypt1p-mediated vesicular transport; cells expressing GTPase-deficient Ypt1-Q67L as the sole Ypt1p have no observable growth, secretion, or morphology defects; GTP hydrolysis is required only for recycling of Ypt1p between compartments. |
Q67L loss-of-GTPase mutant, GAP stimulation assay, secretion assays, electron microscopy |
Molecular and cellular biology |
High |
9447979
|
| 2002 |
Two novel Ypt1-specific GAPs, Gyp5p and Gyp8p, were identified; Gyp5p accelerates Ypt1p GTPase activity 4.2×10⁴-fold; constitutively active Ypt1(Q67L) expressing cells show growth defects and autophagy-like morphology dependent on background of GAP deletion, indicating GTP hydrolysis is needed for balanced ER-Golgi vesicle flow. |
GAP activity biochemical assay, subcellular fractionation, yeast genetics (GAP deletion strains), electron microscopy |
The Journal of biological chemistry |
High |
12189143
|
| 2012 |
Ypt1/Rab1 is recruited to the preautophagosomal structure (PAS) by TRAPPIII (activated via Atg17 recruitment); activated Ypt1 then recruits the Atg1 kinase to the PAS, bringing it near its binding partner Atg17, thereby restricting Atg1 specifically to the PAS for autophagosome formation. |
Yeast two-hybrid, co-immunoprecipitation, fluorescence microscopy, genetic epistasis with atg17 mutants |
Proceedings of the National Academy of Sciences of the United States of America |
High |
23716696
|
| 2012 |
Atg9 vesicles recruit the TRAPPIII complex (via direct Trs85-Atg9 interaction) and Ypt1 to the preautophagosomal structure in an Atg9-dependent manner, establishing a vesicle-tethering machinery for autophagosome formation. |
Atg9 vesicle purification and mass spectrometry, co-IP (Trs85-Atg9), fluorescence microscopy, deletion mutant analysis |
The Journal of biological chemistry |
High |
23129774
|
| 2015 |
Ypt1/Rab1 binds and activates the CK1δ kinase Hrr25 to spatially regulate phosphorylation of the COPII coat, directing ER-derived vesicles to the Golgi; hrr25 mutants are defective in both ER-Golgi trafficking and macroautophagy. |
Co-immunoprecipitation, kinase activity assay, yeast genetic analysis (hrr25 mutants), in vitro transport assay |
The Journal of cell biology |
High |
26195667
|
| 2016 |
TRAPPIII is the primary GEF that activates Ypt1/Rab1 in both Golgi trafficking and autophagy pathways in wild-type yeast; TRAPPII and TRAPPIII are the only two TRAPP complexes present in yeast, contrary to earlier reports of four complexes. |
Biochemical purification of TRAPP complexes, GEF activity assays, genetic analysis, secretion and autophagy assays |
The Journal of cell biology |
High |
29109089
|
| 2016 |
A Trs33-containing TRAPP complex (TRAPP IV) acts as an autophagy-specific GEF for Ypt1; in the absence of Trs85, Trs33 is required for Ypt1-mediated autophagy and recruitment of core-TRAPP and Ypt1 to the PAS. |
Yeast genetics (trs85Δ, trs33Δ double mutants), co-IP, fluorescence microscopy of PAS markers, autophagy assays |
Genetics |
Medium |
27672095
|
| 2010 |
Rab1a regulates cell migration by controlling integrin β1 recycling to lipid rafts and to the plasma membrane; p115 acts as a downstream effector mediating Rab1a regulation of integrin recycling and lipid raft localization. |
RNAi screen, siRNA knockdown, cell adhesion and spreading assays, flow cytometry (lipid raft fractionation), integrin β1 recycling assay, p115 knockdown |
The Journal of biological chemistry |
Medium |
20639577
|
| 2012 |
Rab1A is required for microtubule-dependent anterograde melanosome transport in melanocytes; Rab1A localizes to mature melanosomes; its loss (siRNA or overexpression of dominant-negative TBC1D20 GAP) causes perinuclear melanosome aggregation and specifically suppresses long-range anterograde movements. |
Genome-wide constitutively active/negative Rab screen, siRNA knockdown, immunofluorescence localization, time-lapse live imaging |
Journal of cell science |
High |
22854043
|
| 2015 |
Rab1A recruits SKIP/PLEKHM2 as a specific effector on melanosomes, and Rab1A–SKIP–kinesin-1 (Kif5b/KLC2) form a transport complex that mediates anterograde melanosome transport; Arl8 (not Rab1A) mediates anterograde lysosome transport via the same SKIP-kinesin-1 machinery. |
Co-immunoprecipitation, GST pulldown, siRNA knockdown, live-cell imaging, dominant-negative mutant expression |
Scientific reports |
High |
25649263
|
| 2014 |
Rab1a regulates sorting of early endocytic vesicles for multiple cargo (EGF, transferrin, ASOR); in Rab1a knockdown cells, EGF fails to reach lysosomes and transferrin is redirected to slow Rab11 recycling rather than fast Rab4 recycling; the minus-end-directed kinesin KifC1 is identified as an effector recruited by Rab1a to early endocytic vesicles. |
Stable Rab1a knockdown cell line, fluorescence microscopy tracking of fluorescent cargo, co-immunoprecipitation (KifC1) |
American journal of physiology. Gastrointestinal and liver physiology |
Medium |
24407591
|
| 1996 |
Rab1a is associated not only with ER-to-Golgi transport vesicles but also with postendosomal transcytotic vesicles in rat liver, as demonstrated by immunoisolation of vesicles using anti-Rab1a beads; these vesicles contain transcytotic markers and multiple Rab GTPases. |
Immunoisolation of vesicles using antibody-coated magnetic beads, Western blotting for organelle markers, [α-32P]GTP overlay |
The Journal of biological chemistry |
Medium |
8939959
|
| 2016 |
C9orf72 interacts with Rab1a and the ULK1 autophagy initiation complex; as a Rab1a effector, C9orf72 controls initiation of autophagy by regulating Rab1a-dependent trafficking of the ULK1 complex to the phagophore. |
Co-immunoprecipitation (C9orf72 with Rab1a and ULK1), siRNA knockdown, autophagy flux assays (p62 accumulation, LC3 puncta), iNeuron studies |
The EMBO journal |
High |
27334615
|
| 2018 |
Salmonella effectors SseF and SseG inhibit autophagy by directly interacting with Rab1A, disrupting its interaction with the TRAPPIII GEF complex, thereby blocking Rab1A activation, ULK1 recruitment, and phosphatidylinositol 3-phosphate biogenesis, ultimately impairing autophagosome formation. |
Co-immunoprecipitation (SseF/SseG with Rab1A and TRAPPIII), GEF disruption assay, ULK1 recruitment assay, PI3P measurement, siRNA rescue experiments, mouse infection model |
The Journal of biological chemistry |
High |
29610274
|
| 2021 |
M. tuberculosis PE_PGRS20 and PE_PGRS47 inhibit autophagy initiation by directly interacting with Rab1A; silencing Rab1A rescues the survival defects of pe_pgrs20/pe_pgrs47 deletion mutants, placing Rab1A as the host target of these mycobacterial effectors. |
Co-immunoprecipitation (PE_PGRS proteins with Rab1A), Rab1A silencing rescue, autophagy flux assays in macrophages |
mSphere |
Medium |
34346699
|
| 2017 |
Rab1a interacts with and regulates optineurin (OPTN)-induced autophagosome formation in neuroblastoma cells; a zinc finger domain of OPTN is required for Rab1a binding; active Rab1a (GTP-bound) is required for the interaction; Rab1a knockdown inhibits OPTN-induced LC3-EGFP translocation to autophagosomes. |
Yeast two-hybrid, co-immunoprecipitation, confocal microscopy colocalization, dominant-negative/constitutively-active Rab1a mutants, LC3-EGFP autophagosome translocation assay |
Journal of neuroscience research |
Medium |
28843006
|
| 2014 |
Hsc70 interacts with Rab1A in a chaperone-dependent manner and prevents its ubiquitination and degradation under proteotoxic stress; Hsc70 knockdown decreases Rab1A levels and increases its ubiquitination; Rab1A knockdown inhibits autophagosome formation and induces cell death under stress. |
Anti-Hsc70 affinity purification + mass spectrometry, co-immunoprecipitation, ubiquitination assay, siRNA knockdown, autophagy assay |
PloS one |
Medium |
24801886
|
| 2022 |
The E3 ubiquitin ligase RNF115 catalyzes K11-linked ubiquitination of RAB1A at Lys49 and Lys61; this modification impairs recruitment of GDI1 to RAB1A, preventing its reactivation cycle; RAB1A ubiquitination-resistant mutant (K49/61R) promotes ER-to-Golgi trafficking of TLRs. |
Co-immunoprecipitation (RNF115 with RAB1A), in vitro ubiquitination assay, site-directed mutagenesis (K49R/K61R), GDI1 recruitment assay, TLR trafficking assay in Rnf115+/+ and Rnf115−/− cells |
Advanced science |
High |
35343654
|
| 2020 |
Ubiquitin-specific protease USP2a deubiquitinates and stabilizes RAB1A, preventing its proteasomal degradation; this stabilization is required for USP2a-mediated HCC progression. |
LC-MS/MS affinity proteomics, co-immunoprecipitation, ubiquitination assay, siRNA knockdown, in vitro and in vivo tumor models |
Cellular oncology |
Medium |
33074477
|
| 2024 |
During mitosis, Aurora kinase A (AURKA) phosphorylates Rab1A at Thr75; structural analysis shows Thr75 phosphorylation locks Rab1A in a constitutively active state by preventing GDI interaction; activated Rab1A is retained on the ER and induces oligomerization of RTN and REEP ER-shaping proteins, increasing ER complexity; this mechanism is evolutionarily conserved from C. elegans and Drosophila to mammals. |
Structural analysis (crystal/cryo-EM implied), in vitro kinase assay (AURKA phosphorylation of Rab1A), site-directed mutagenesis (T75A), GDI interaction assay, ER morphology assay, genetic modification in C. elegans/Drosophila/mammalian cells |
Nature structural & molecular biology |
High |
38177680
|
| 2011 |
In Dictyostelium, Rab1A binds the LRRK2-related Roco kinase Roco2 in vivo; active GTP-bound Rab1A regulates Roco2 kinase activity; Roco2 in turn controls F-actin polymerization and pseudopod extension by acting upstream of filamin (abp120), establishing a Rab1A-Roco2-filamin pathway regulating actin cytoskeleton and cell motility. |
Co-immunoprecipitation (Roco2 with Rab1A and filamin), kinase activity assay, constitutively active/dominant-negative Rab1A mutants, chemotaxis assays, F-actin measurement |
Molecular biology of the cell |
Medium |
21551065
|
| 2011 |
Rab1A (and Rab43) is required for herpes simplex virus 1 secondary envelopment; depletion of Rab1a/b via their specific GAP TBC1D20 prevents viral glycoproteins from trafficking from the ER to the viral assembly compartment, resulting in accumulation of unenveloped particles. |
Overexpression of 37 Rab-GAPs (functional screen), siRNA knockdown, infectious titer assay, immunofluorescence microscopy of viral glycoproteins, electron microscopy |
Journal of virology |
Medium |
21680502
|
| 2014 |
RAB1A is required for Vaccinia virus production of intracellular enveloped virions (IEVs) but not intracellular mature virions (IMVs); RAB1A facilitates the wrapping step consistent with its role in ER-to-Golgi transport. |
siRNA knockdown, infectious titer assay, immunofluorescence, electron microscopy |
Virology |
Medium |
25462347
|
| 2017 |
Rab1A is required for classical swine fever virus (CSFV) particle assembly; Rab1A binds CSFV NS5A protein; Rab1A depletion reduces intracellular and extracellular viral titers without affecting viral genome replication or E2 protein expression, specifically implicating particle assembly. |
siRNA knockdown, viral titer assay, qRT-PCR (genome copies), co-immunoprecipitation (Rab1A–NS5A), confocal microscopy, neutralizing antibody blocking assay |
Virology |
Medium |
29128753
|
| 2011 |
Rab1A overexpression prevents Golgi fragmentation caused by alpha-synuclein in dopaminergic neurons; the non-prenylable Rab1A-ΔCC mutant fails to rescue Golgi morphology, linking the protective effect to Rab1A's ER-to-Golgi trafficking activity. |
AAV-mediated co-expression in rat substantia nigra, Golgi morphology assay, non-prenylable mutant analysis, motor behavior assay |
Journal of Parkinson's disease |
Medium |
23939344
|
| 2012 |
Ypt1 protein specifically associates in vivo with unspliced HAC1 RNA and promotes HAC1 RNA decay; this association is disrupted during ER stress induction of the UPR; Ypt1 depletion reduces HAC1 RNA decay rate, elevating HAC1 levels and delaying UPR attenuation. |
Protein microarray screening for HAC1 RNA binding, RNA co-immunoprecipitation, HAC1 RNA decay assays, genetic analysis (ire1Δ, ada5Δ) |
PLoS genetics |
Medium |
22844259
|
| 2020 |
Human GOLPH3 is a direct effector of RAB1A and RAB1B; interaction is nucleotide-dependent, favored by GTP-locked (active) variants; expression of GTP-locked RAB1A reduces GOLPH3 localization at the Golgi, suggesting Rab1A-GTP releases GOLPH3 from the Golgi. |
Co-immunoprecipitation (GOLPH3 with RAB1A/B wild-type and GTP-locked mutants), fluorescence microscopy of GOLPH3 distribution |
PloS one |
Medium |
32790781
|
| 2021 |
Rab1A mediates amino acid (particularly branched-chain amino acid)-dependent mTORC1 signaling in vivo; whole-body Rab1A knockout mice are hyperglycemic and glucose intolerant due to impaired insulin transcription and β-cell proliferation/maintenance; mechanistically, Rab1A mediates BCAA-mTORC1 signaling to regulate stability and localization of the insulin transcription factor Pdx1. |
Tamoxifen-induced whole-body Rab1A knockout mouse, glucose tolerance test, insulin assay, β-cell analysis, Pdx1 localization and stability assay |
Cell reports |
High |
33730578
|
| 2021 |
NAB2 (an N-arylbenzimidazole small molecule) binds Rab1a selectively in its GDP-bound form; NAB2 binding phenocopies Rab1a overexpression in alleviating α-synuclein toxicity; Rab1a was identified as a target of NAB2 by Thermal Proteome Profiling and SPROX chemoproteomic analyses. |
Thermal Proteome Profiling (TPP), SPROX chemoproteomic analysis, cellular phenotypic assay (α-synuclein toxicity rescue), GDP/GTP-state selectivity binding assay |
RSC chemical biology |
Medium |
35128413
|
| 2022 |
Rab1a interacts with ULK1 and promotes ULK1 phosphorylation dependent on its GTP-binding activity; this Rab1a-ULK1 interaction initiates autophagy that promotes PRRSV replication. |
Co-immunoprecipitation (Rab1a with ULK1), ULK1 phosphorylation assay, GTP-binding mutant analysis, ATG7 knockdown rescue, viral titer assay |
Virus research |
Medium |
36306941
|
| 2005 |
Yeast Yip3p (a GDI displacement factor ortholog) forms a distinct complex with Ypt1p that is separable from its complex with reticulon Rtn1p; Yip3p is predominantly Golgi-localized; however, loss of Yip3p or Rtn1p does not perturb intracellular Rab localization, indicating Yip3p-Ypt1p interaction is not critical for Rab membrane targeting. |
Co-purification from detergent extracts, Western blotting, fluorescence microscopy in yip3Δ and rtn1Δ mutants |
Eukaryotic cell |
Medium |
16002643
|
| 1998 |
A Ypt1p-specific GEF activity localizes to the acceptor (Golgi) fraction rather than the donor (ER) fraction; a Ypt1p-specific GAP activity colocalizes with ER markers; the GEF is specific for Ypt1p and not other Ypt proteins; the GAP has higher affinity for GTP-bound Ypt1p. |
Partial purification of GEF and GAP activities from yeast fractions, GDP-release and GTP-uptake assays, GTPase stimulation assay, subcellular fractionation |
Molecular biology of the cell |
Medium |
9763446
|