| 1992 |
SEC6 encodes an 85 kDa predominantly soluble protein that sediments at 14S and is required for fusion of post-Golgi vesicles with the plasma membrane in S. cerevisiae; sec6-4 displays synthetic lethality with sec8-9, indicating inter-related functions between the two gene products. |
Gene cloning by complementation, subcellular fractionation, gene disruption, synthetic lethality analysis |
Yeast |
High |
1523887
|
| 1995 |
Sec6 is a stable component of the yeast Sec6/8/15 multisubunit complex (~1-2 MDa); Sec6 co-fractionates with Sec8/15 by metal-affinity chromatography, gel filtration, and sucrose velocity centrifugation, and coimmunoprecipitates with c-myc-tagged Sec8. The complex is disrupted in sec3-2, sec5-24, and sec10-2 backgrounds. Sec8 localizes to small bud tips, placing the complex at sites of exocytosis. |
Immobilized metal affinity chromatography, gel filtration, sucrose velocity centrifugation, immunoprecipitation, immunofluorescence localization |
The Journal of cell biology |
High |
7615633
|
| 1998 |
In mammalian epithelial (MDCK) cells, the Sec6/8 complex resides in a cytosolic ~17S complex; upon calcium-dependent cell-cell adhesion, ~70% of Sec6/8 is recruited to sites of cell-cell contact. Sec8 antibodies in permeabilized cells inhibit delivery of LDL receptor to the basal-lateral membrane but not p75NTR to the apical membrane, demonstrating that lateral membrane recruitment of the Sec6/8 complex is essential for biogenesis of epithelial cell surface polarity. |
Sucrose gradient sedimentation, streptolysin-O permeabilization with antibody inhibition, immunofluorescence |
Cell |
High |
9630218
|
| 1998 |
The rat brain sec6/8 complex coimmunoprecipitates with a filament composed of four mammalian septins (including CDC10), suggesting a physical interaction between the exocyst and septin filaments. Electron microscopy of glutaraldehyde-fixed rat brain sec6/8 complex reveals a T- or Y-shaped conformation. |
Co-immunoprecipitation, electron microscopy, sucrose gradient fractionation |
Neuron |
Medium |
9655500
|
| 2001 |
Sec6/8 complex is present on both the trans-Golgi network (TGN) and plasma membrane in NRK cells, colocalizing with exocytic cargo VSVG-tsO45. Brefeldin A blocks Sec6/8 recruitment to the plasma membrane; expression of kinase-inactive protein kinase D or low-temperature incubation causes Sec6/8 accumulation on TGN. Antibodies against TGN-bound or plasma membrane-bound Sec6/8 added to semiintact cells cause cargo accumulation in respective compartments, indicating Sec6/8 is required for multiple steps of TGN-to-plasma membrane exocytic transport. |
Immunofluorescence colocalization, semiintact cell functional assays with specific antibody inhibition, pharmacological treatments |
The Journal of cell biology |
High |
11696560
|
| 2001 |
Human Sec3 (hSec3) interacts with Sec5 and Sec8 subunits of the mammalian Sec6/8 complex in the yeast two-hybrid system. GFP-fusions of most subunits fail to assemble into complex with endogenous proteins and are cytosolic when expressed in MDCK cells; only GFP-Exo70 localizes to lateral membrane, and its overexpression disrupts tight monolayer formation. |
Yeast two-hybrid, GFP fusion expression and imaging in MDCK cells |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
11493706
|
| 2000 |
Sec6/8 complex associates with Ca2+ signaling proteins at the apical pole of pancreatic acinar cells. Immunoprecipitation of Sec8 co-precipitates Sec6, IP3R3, Gβγ, plasma membrane Ca2+ pump, Gαq, PLCβ1, and IP3R1. This interaction is mediated by the actin cytoskeleton, as actin filament disruption dissociates Sec6/8 from Ca2+ signaling proteins. Anti-Sec6/8 antibodies inhibit Ca2+ signaling upstream of Ca2+ release by IP3; actin disruption by latrunculin B partially translocates Sec6/8 to cytosol and interferes with Ca2+ wave propagation. |
Immunoprecipitation, confocal immunolocalization, pharmacological actin disruption, functional Ca2+ signaling assays |
The Journal of cell biology |
Medium |
10973998
|
| 2003 |
Crystal structure of the Sec5 Ral-binding domain in complex with RalA·GppNHp at 2.1 Å resolution shows that Sec5 folds into an immunoglobulin-like β-sandwich and interacts with RalA via a continuous antiparallel β-sheet involving both switch regions. Sec5 Thr11 and Arg27, and RalA Glu38 are required for complex formation (validated by isothermal titration calorimetry). This establishes the structural basis for GTP-dependent RalA binding to the Sec6/8 complex via the Sec5 subunit. |
X-ray crystallography (2.1 Å), isothermal titration calorimetry, site-directed mutagenesis |
The EMBO journal |
High |
12839989
|
| 2004 |
In polarized MDCK epithelial cells, the Sec6/8 complex is recruited to cell-cell contacts in a high molecular mass complex with tight junction proteins and a portion of E-cadherin. Sec6/8 co-immunoprecipitates with cell surface-labeled E-cadherin and nectin-2α. Co-expression of E-cadherin and nectin-2α in fibroblasts is sufficient to recruit Sec6/8 to cell-cell contacts, indicating that adhesion complexes specify Sec6/8 localization. |
Co-immunoprecipitation, sucrose gradient fractionation, GFP overexpression in fibroblasts, immunofluorescence |
Journal of cell science |
Medium |
14709721
|
| 2005 |
Crystal structure of the Exo84 Ral-binding domain in complex with active RalA shows the domain adopts a pleckstrin homology fold. Structural and biochemical data demonstrate that Exo84 and Sec5 competitively bind active RalA via an overlapping interface including both switch regions; key binding residues were confirmed by mutagenesis. This establishes that RalA regulates Sec6/8 complex assembly through competitive effector binding. |
X-ray crystallography, site-directed mutagenesis, binding assays |
The EMBO journal |
High |
15920473
|
| 2005 |
In Drosophila photoreceptor cells (PRCs), loss-of-function sec6 mutations cause cell lethality, disrupt plasma membrane growth, and lead to accumulation of secretory vesicles and failure to transport proteins to the rhabdomere (apical subdomain). Sec6 but not Sec5 or Sec8 shows accumulation at adherens junctions in developing PRCs. Rab11 forms a complex with Sec5, and Sec5 interacts with Sec6, suggesting the exocyst is a Rab11 effector for apical membrane protein transport. |
Genetic loss-of-function analysis, immunofluorescence localization, co-immunoprecipitation |
The Journal of cell biology |
High |
15897260
|
| 2005 |
In Drosophila epithelial cells, loss of function of exocyst components sec5, sec6, and sec15 causes DE-Cadherin accumulation in an enlarged Rab11 recycling endosomal compartment and inhibits DE-Cad delivery to the plasma membrane. Rab11 and Armadillo interact with exocyst components Sec15 and Sec10 respectively, placing Sec6-containing exocyst complex at the step of Rab11 recycling endosome-to-membrane trafficking. |
Genetic loss-of-function, immunofluorescence, co-immunoprecipitation |
Developmental cell |
High |
16224820
|
| 2005 |
The sec6-4 phenotype in S. cerevisiae is defined by a single point mutation L633P in the SEC6 coding region. At restrictive temperature, Sec6-4p is mislocalized and cells accumulate homogeneous secretory vesicles. At permissive temperature, wild-type Sec6p-GFP localizes to buds and septa, consistent with its role at sites of exocytosis. |
Site-directed mutagenesis, GFP fusion localization, electron microscopy of vesicle accumulation |
Gene |
Medium |
16185821
|
| 2011 |
Yeast Sec6 directly binds Sec1 (a Sec1/Munc18 family SM protein) in addition to its known binding partner Sec9 (plasma membrane SNARE). The Sec6-Sec1 interaction is exclusive of Sec6-Sec9 but compatible with Sec6-exocyst assembly. The Sec6-exocyst interaction is incompatible with Sec6-Sec9. This proposes a sequential mechanism whereby vesicle arrival triggers Sec6 to release Sec9, assemble into exocyst, and recruit Sec1 for coordinated SNARE complex formation. |
In vitro binding assays, co-immunoprecipitation, yeast genetics |
Molecular biology of the cell |
High |
22114349
|
| 2015 |
S. cerevisiae Sec6 binds both the binary Sso1-Sec9 and ternary Sso1-Sec9-Snc2 SNARE complexes; it does not change the rate of SNARE assembly (contrary to previous hypothesis that it inhibits assembly). Cross-linking/mass spectrometry identified specific residues required for Sec6-Sec9 binding; mutation of these residues causes a yeast growth defect. |
In vitro SNARE assembly kinetic assays, cross-linking mass spectrometry, site-directed mutagenesis, yeast growth assays |
The Journal of biological chemistry |
High |
26446795
|
| 2012 |
Sec6 knockdown in HSC3 oral cancer cells increases α-E-catenin expression and causes E-cadherin and β-catenin to localize predominantly at cell-cell contact regions, indicating Sec6 negatively regulates cell-cell adhesion complex organization. |
siRNA knockdown, immunofluorescence, western blot |
FEBS letters |
Low |
22381337
|
| 2014 |
Sec6 regulates cytoplasmic translocation and degradation of p27 via interactions with Jab1/CSN5 and Siah1; Sec6 promotes p27 phosphorylation at Thr157, facilitating cytoplasmic localization and subsequent degradation, thereby suppressing cell cycle progression. |
siRNA knockdown, co-immunoprecipitation, phosphorylation analysis, cell cycle assays |
Cellular signalling |
Medium |
24949832
|
| 2015 |
Perturbation of exocyst components including Sec6/8 results in resistance to ionizing radiation and accelerated resolution of DNA damage, but with accumulation of aberrant chromatid exchanges. Sec8 perturbation leads to accumulation of ATF2, RNF20, and DDR-associated chromatin marks, and Rad51 repairosomes, indicating exocyst supports DNA repair fidelity. |
siRNA knockdown, irradiation survival assays, chromatin immunoprecipitation, recombination frequency measurement |
Molecular and cellular biology |
Medium |
26283729
|
| 2016 |
Sec6 knockdown inhibits IκBα degradation, delays nuclear translocation of p65, and reduces NF-κB transcriptional activity in TNF-α-stimulated HeLa cells. Mechanistically, Sec6 knockdown decreases expression of p90RSKs and phosphorylation of ERK, p90RSK1 (Ser380), and IκBα (Ser32). |
siRNA knockdown, western blot for phosphorylation, luciferase transcriptional reporter, immunofluorescence for p65 localization |
Journal of cellular physiology |
Medium |
26247921
|
| 2018 |
Sec6 knockdown suppresses phosphorylation of p38 MAPK (via MKK3/6), MK2, and HSP27 (at Ser78 and Ser82), and overexpression has the reverse effect. Reduced phospho-HSP27 via Sec6 knockdown suppresses cell migration and promotes apoptosis after TNF-α/cycloheximide treatment. |
siRNA knockdown, overexpression, western blot for kinase phosphorylation, cell migration assays, apoptosis assays |
Cellular signalling |
Medium |
29729335
|
| 2019 |
In Physcomitrella patens, exocyst subunit Sec6 (but not Sec3 or Sec5) localizes to microtubule overlap regions in the phragmoplast prior to cell plate membrane arrival. Sec6 physically interacts with KEULE (a Sec1/Munc18 ortholog). Sec6 gene silencing delays recruitment of KEULE and vesicles to the early cell plate, indicating Sec6 promotes vesicle-vesicle fusion at microtubule overlaps independently of vesicle delivery. |
Live-cell fluorescence microscopy, co-immunoprecipitation, gene silencing, time-lapse imaging |
Journal of cell science |
Medium |
30635445
|
| 2020 |
Tnfaip2/Exoc3 acts epistatically upstream of vimentin (Vim) in controlling lipid metabolism during stem cell differentiation; Tnfaip2 knockout impairs induction of triacylglycerol synthesis and lipid droplet formation in differentiating ESCs. Knockdown of planarian Smed-exoc3 also causes strong reduction of TAGs. Supplementation with palmitic acid rescues both ESC differentiation and planarian organ maintenance, placing Exoc3 upstream of TAG/lipid droplet biosynthesis required for differentiation. |
Genetic knockout/knockdown, lipidomics, epistasis analysis, rescue with palmitic acid supplementation |
EMBO reports |
Medium |
33300287
|
| 2021 |
EXOC3 conditional knockout in mouse megakaryocytes/platelets causes defects in platelet aggregation, integrin activation, α-granule secretion (P-selectin, PF4), dense granule secretion, and lysosomal granule secretion after GPVI agonist stimulation. GPVI surface levels were decreased 14.5% in KO platelets, with defects in proximal GPVI signaling (Syk and LAT phosphorylation) and calcium mobilization. PAR4-stimulated responses were paradoxically enhanced in KO platelets, and suppressed by P2Y12 antagonist, implicating enhanced ADP release. EXOC3 KO mice show accelerated arterial thrombosis and improved hemostasis. |
Conditional knockout mice, platelet aggregometry, granule secretion assays, flow cytometry, phosphorylation analysis, calcium imaging, in vivo thrombosis model, tail bleeding time |
Blood advances |
High |
33560379
|
| 2023 |
Two truncated forms of human Sec6 (HuSec6 121-734 and HuSec6 121-745) were expressed, purified (>95% purity), and crystallized; X-ray diffraction yielded ~9 Å resolution crystals, providing initial structural data for human Sec6. |
Recombinant protein expression in E. coli, X-ray crystallography (9 Å resolution) |
Studies in health technology and informatics |
Low |
38007759
|
| 2024 |
Male germ cell-specific conditional knockout of Exoc3 (SEC6) in mice does not cause spermatogenesis defects, establishing that EXOC3 is not required for spermatogenesis (negative finding). |
Conditional knockout mice, histological analysis of spermatogenesis |
Experimental animals |
Medium |
38325858
|
| 2025 |
Sec6 suppresses innate immune responses to bovine herpesvirus 1 (BoHV-1) by promoting NDP52-mediated autophagic degradation of STING. Mechanistically, Sec6 enhances the physical interaction between NDP52 and STING; NDP52 knockdown abolishes Sec6-mediated IFN-β suppression and Sec6's ability to enhance viral replication. |
Overexpression/knockdown, co-immunoprecipitation, autophagic flux assays, IFN-β reporter assay, viral replication assay |
Veterinary microbiology |
Medium |
41406560
|
| 2025 |
Sec6 suppresses BEFV-triggered type I IFN signaling by promoting P62-mediated autophagic degradation of MAVS. Sec6 enhances the P62-MAVS interaction; P62 knockdown abolishes Sec6-mediated IFN-I suppression and viral replication enhancement. Sec6 fails to degrade MAVS in P62-knockdown cells. |
Overexpression/knockdown, co-immunoprecipitation, autophagic flux assays, IFN-I reporter assay, viral replication assay |
Veterinary microbiology |
Medium |
40780029
|