CoChat
Sign Up
Cited by 5Cellular transport and secretionAutophagy in Disease and TherapyEndoplasmic Reticulum Stress and Disease

The DOI matches its Crossref record; 2 of the 2 references it cites were found in a registry or an index.

TUG protein acts through a disordered region to organize the early secretory pathway

Open access
Parchure, Anup, Tejada, Helen, Xi, Zhiqun, Kim, Yeongho, Su, Maohan, Yan, You, Julca-Zevallos, Omar, Alcázar-Román, Abel R., Villemeur, Marie, Liu, Xinran, Toomre, Derek, Raote, Ishier, Bogan, Jonathan S.·2025·Nature Communications
DOI 10.1038/s41467-025-60691-8PMC PMC12218103

Published version of a preprint

Open this paper in CoChat — free accountOpen PDF

Abstract

The Endoplasmic Reticulum (ER)-Golgi Intermediate Compartment (ERGIC) is a network of tubules and vesicles known for producing COPI vesicles and receiving COPII vesicles from the ER. Much about its identity, stability, and regulation remains unknown. Here, we show that TUG (UBXN9, Aspscr1) protein, a central regulator of GLUT4 trafficking, localizes to the ERGIC, and that its deletion enhances anterograde flux of a model soluble cargo protein. TUG deletion redistributes ERGIC markers to the cis-Golgi and alters Golgi morphology. TUG forms biomolecular condensates in vitro and contains a central disordered region that mediates its recruitment to ERGIC membranes. A distinct N-terminal region mediates its oligomerization in cells. TUG deletion disrupts ERGIC-dependent processes, including autophagy and collagen secretion, and alters the targeting of the CFTR chloride channel. We conclude that TUG organizes and stabilizes ERGIC membranes to support their roles in diverse secretory and degradative membrane trafficking pathways.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1038/s41467-025-60691-8

Cites

2 of 2 references verified0 citation errors · 0 not checkable

Statistics

Figures

Fig. 1 TUG localizes to the ERGIC. a–d Confocal images are shown of TUG KO HeLa cells co-transfected with mCherry-tagged TUG (Tug-mCherry; magenta) together with Emerald-tagged ERGIC53 (Em-ERGIC53; a; green), the ERES marker Sec23-GFP (b; green), the Golgi marker mNG-tagged GMAP210 (GMAP210-mNG; c; green) and Stx12-GFP (d; green). Note the colocalization between TUG and ERGIC53 in peripheral punctate structures. TUG puncta also are positive for Stx12 (yellow circles), but are distinct from the ERES (Sec23) and are excluded from the Golgi apparatus (GMAP210). Images for co-transfection of Tug-mCherry with Em-ERGIC53 are representative of three independent experiments with similar results. Those for Sec23-GFP and GMAP210-mNG and Stx12-GFP are representative of two independent experiments with similar results.
Fig. 1 TUG localizes to the ERGIC. a–d Confocal images are shown of TUG KO HeLa cells co-transfected with mCherry-tagged TUG (Tug-mCherry; magenta) together with Emerald-tagged ERGIC53 (Em-ERGIC53; a; green), the ERES marker Sec23-GFP (b; green), the Golgi marker mNG-tagged GMAP210 (GMAP210-mNG; c; green) and Stx12-GFP (d; green). Note the colocalization between TUG and ERGIC53 in peripheral punctate structures. TUG puncta also are positive for Stx12 (yellow circles), but are distinct from the ERES (Sec23) and are excluded from the Golgi apparatus (GMAP210). Images for co-transfection of Tug-mCherry with Em-ERGIC53 are representative of three independent experiments with similar results. Those for Sec23-GFP and GMAP210-mNG and Stx12-GFP are representative of two independent experiments with similar results.CC BY · PubMed Central
Fig. 2 TUG regulates anterograde trafficking. a WT (top) and TUG KO (bottom) MEFs were fixed and stained using antibodies to the ERGIC marker ERGIC53 (magenta) and the cis-Golgin GM130 (green). Images are a single slice from a confocal stack. Dashed boxes in the top image denote peripheral punctate staining of ERGIC53, which is devoid of GM130. The dashed region in the bottom image denotes the Golgi, based on GM130 staining. Note the reduction in peripheral ERGIC53 puncta in TUG KO MEFs, compared to WT cells, and increased ERGIC53 distribution at the cis-Golgi. b Confocal images were segmented to generate ERGIC53 and GM130 surfaces. The graph compares independent ERGIC53 surfaces (not touching GM130 surfaces) in WT and TUG KO MEFs. A reduction in the number of independent ERGIC53 surfaces was observed in TUG KO MEFs. Data from 16 WT and 16 KO cells are shown; mean ± standard deviation (s.d.). c The graph compares average intensities of ERGIC53 staining at the Golgi after collapsing a confocal stack. Data from 16 WT and 16 KO cells are shown; mean ± s.d. d Images of TUG KO (top) and WT (bottom) MEFs containing retroviruses to express PAUF tagged with mKate2 and FM4 domains. The FM4 domains cause aggregation of PAUF in the ER (marked by PDI, in green; left), and is distinct from GM130 (green; right) signal in the absence of D/D solubilizer. The addition of D/D-solubilizer releases PAUF from the ER for anterograde traffic through the secretory pathway. Cells were fixed 10 min after the addition of D/D solubilizer and were imaged (right). In TUG KO MEFs (top), most signal is concentrated at the Golgi, but in WT MEFs there is also surrounding ER signal. e Data from replicates of the experiment in f were quantified, and enrichment of mKate2-tagged PAUF signal at the Golgi is normalized to the signal in the surrounding ER. N = 42 cells in each group, acquired from 4 independent experiments. The box indicates median and quartile ranges, and whiskers show the spread of the data from minimum to maximum. All statistical analyses used an unpaired two-tailed t test with Welch’s correction.
Fig. 2 TUG regulates anterograde trafficking. a WT (top) and TUG KO (bottom) MEFs were fixed and stained using antibodies to the ERGIC marker ERGIC53 (magenta) and the cis-Golgin GM130 (green). Images are a single slice from a confocal stack. Dashed boxes in the top image denote peripheral punctate staining of ERGIC53, which is devoid of GM130. The dashed region in the bottom image denotes the Golgi, based on GM130 staining. Note the reduction in peripheral ERGIC53 puncta in TUG KO MEFs, compared to WT cells, and increased ERGIC53 distribution at the cis-Golgi. b Confocal images were segmented to generate ERGIC53 and GM130 surfaces. The graph compares independent ERGIC53 surfaces (not touching GM130 surfaces) in WT and TUG KO MEFs. A reduction in the number of independent ERGIC53 surfaces was observed in TUG KO MEFs. Data from 16 WT and 16 KO cells are shown; mean ± standard deviation (s.d.). c The graph compares average intensities of ERGIC53 staining at the Golgi after collapsing a confocal stack. Data from 16 WT and 16 KO cells are shown; mean ± s.d. d Images of TUG KO (top) and WT (bottom) MEFs containing retroviruses to express PAUF tagged with mKate2 and FM4 domains. The FM4 domains cause aggregation of PAUF in the ER (marked by PDI, in green; left), and is distinct from GM130 (green; right) signal in the absence of D/D solubilizer. The addition of D/D-solubilizer releases PAUF from the ER for anterograde traffic through the secretory pathway. Cells were fixed 10 min after the addition of D/D solubilizer and were imaged (right). In TUG KO MEFs (top), most signal is concentrated at the Golgi, but in WT MEFs there is also surrounding ER signal. e Data from replicates of the experiment in f were quantified, and enrichment of mKate2-tagged PAUF signal at the Golgi is normalized to the signal in the surrounding ER. N = 42 cells in each group, acquired from 4 independent experiments. The box indicates median and quartile ranges, and whiskers show the spread of the data from minimum to maximum. All statistical analyses used an unpaired two-tailed t test with Welch’s correction.CC BY · PubMed Central
Fig. 3 TUG regulates the organization of the Golgi apparatus. a WT and TUG KO MEFs were immunostained to detect KDELr and GM130. Note the compacted Golgi morphology in TUG KO MEFs. b Quantification of Golgi length along the nuclear circumference. Plots show mean ± s.d.; N = 35 WT and 33 KO cells, analyzed using a two-tailed t test. c Compaction of Golgi based on GM130 images was calculated using a circularity function (see Methods). Plots show mean ± s.d.; N = 24 WT and 29 KO cells, analyzed using a two-tailed t test. d Electron microscopy (EM) tomography imaging of Golgi complexes in WT and TUG KO MEFs. At left, cross sections of TUG KO (upper) and WT (lower panels) MEFs are shown. Narrow, well-stacked cisternae are pseudo-colored in blue, dilated cisternae are salmon, and ballooned areas are marked by asterisks. At right, tomographic reconstructions of Golgi complexes in TUG KO and WT MEFs are shown. In addition to dilated cisterna, accumulated small vesicles were observed in TUG KO cells (arrows). e 4Pi-SMS side view images of Golgi stacks are shown, immunostained for GM130 (cis-Golgi; magenta) and Golgin97 (trans-Golgi; green). Images from WT and TUG KO MEFs are indicated. Shaded areas were used to quantify the separation of cis and trans markers in line scan profiles. f Line scan profiles were generated for GM130 and Golgin97. The intensity profile in each channel was fitted to a gaussian and the peak-to-peak distance was used to measure the separation between these cis and trans markers. In the example shown, this distance was 264 nm in WT MEFs and 323 nm in TUG KO MEFs; the increase in KO cells was ~60 nm. FWHM represents full width at half-maximum. g Violin plots of the shortest distances of trans points to the cis isosurface for four WT and four TUG KO cells (see Supplementary fig. 4). The median value for each cell is indicated. For WT cells, N = 254,025, 291,063, 372,879, and 252,424 measurements; for KO cells, N = 290,987, 272,770, 156,019, and 163,102 measurements. The overall median distance was 155.0 nm for WT cells and 210.2 nm for KO cells.
Fig. 3 TUG regulates the organization of the Golgi apparatus. a WT and TUG KO MEFs were immunostained to detect KDELr and GM130. Note the compacted Golgi morphology in TUG KO MEFs. b Quantification of Golgi length along the nuclear circumference. Plots show mean ± s.d.; N = 35 WT and 33 KO cells, analyzed using a two-tailed t test. c Compaction of Golgi based on GM130 images was calculated using a circularity function (see Methods). Plots show mean ± s.d.; N = 24 WT and 29 KO cells, analyzed using a two-tailed t test. d Electron microscopy (EM) tomography imaging of Golgi complexes in WT and TUG KO MEFs. At left, cross sections of TUG KO (upper) and WT (lower panels) MEFs are shown. Narrow, well-stacked cisternae are pseudo-colored in blue, dilated cisternae are salmon, and ballooned areas are marked by asterisks. At right, tomographic reconstructions of Golgi complexes in TUG KO and WT MEFs are shown. In addition to dilated cisterna, accumulated small vesicles were observed in TUG KO cells (arrows). e 4Pi-SMS side view images of Golgi stacks are shown, immunostained for GM130 (cis-Golgi; magenta) and Golgin97 (trans-Golgi; green). Images from WT and TUG KO MEFs are indicated. Shaded areas were used to quantify the separation of cis and trans markers in line scan profiles. f Line scan profiles were generated for GM130 and Golgin97. The intensity profile in each channel was fitted to a gaussian and the peak-to-peak distance was used to measure the separation between these cis and trans markers. In the example shown, this distance was 264 nm in WT MEFs and 323 nm in TUG KO MEFs; the increase in KO cells was ~60 nm. FWHM represents full width at half-maximum. g Violin plots of the shortest distances of trans points to the cis isosurface for four WT and four TUG KO cells (see Supplementary fig. 4). The median value for each cell is indicated. For WT cells, N = 254,025, 291,063, 372,879, and 252,424 measurements; for KO cells, N = 290,987, 272,770, 156,019, and 163,102 measurements. The overall median distance was 155.0 nm for WT cells and 210.2 nm for KO cells.CC BY · PubMed Central
Fig. 4 The central disordered region (IDR1) in TUG is necessary and sufficient to mediate TUG localization to the ERGIC. a Schematic of TUG variants used for expression in TUG KO HeLa cells. All the proteins also contained a C-terminal mCherry tag to aid the visualization of the proteins in living cells. b Images of TUG KO HeLa cells transfected with full-length TUG (Tug-mCherry; left) or with a variant where the central IDR, IDR1 was deleted (Tug-IDR1∆-mCherry; right). Although full-length TUG forms punctate structures, the mutant is present in a diffuse distribution. Also note that the mutant is excluded from the nucleus. c Images of TUG KO HeLa cells transfected with IDR1 alone, tagged with mCherry (IDR1-mCherry; left), or with IDR2 alone, also tagged with mCherry (IDR2-mCherry; right). Although IDR1 forms punctate structures in cells, IDR2 is present in a diffuse distribution. In (b) and (c), a minimum of three independent transfections were performed for Tug-mCherry and IDR1-mCherry, three independent transfections for IDR2-mCherry and two independent transfections for Tug-IDR1∆-mCherry. d Images of TUG KO HeLa cells transfected with IDR1-mCherry and treated with 1,6-hexanediol and imaged before (left) or 150 sec. after 1,6-hexanediol treatment (right). Note that the punctate structures containing IDR1-mCherry continue to be present after 1,6-hexanediol treatment. The structures before and after are not necessarily at the same position, due to shift and refocusing during imaging after drug treatment. The treatment with 1,6-hexanediol was done three times, with similar results each time. e Images of TUG KO HeLa cells transfected with IDR1-mCherry (left) and mCherry-tagged IDR from FUS protein (right). Note the differences in the distribution of the two IDRs in cells. mCherry-tagged FUS IDR was transfected in TUG KO HeLa cells in two independent experiments, with similar results. f Images from TUG KO HeLa cells co-transfected with IDR1-mCherry (red) and Em-ERGIC53 (green). Note the colocalization of the two proteins in peripheral punctate structures. g Images from TUG KO HeLa cells co-transfected with IDR1-mCherry (red) and Stx12-GFP (green). Note the partial colocalization of the two proteins in punctate structures. Representative images for colocalization of IDR1-mCherry were from three independent experiments with Em-ERGIC53 (f) and two independent experiments with Stx12-GFP (g).
Fig. 4 The central disordered region (IDR1) in TUG is necessary and sufficient to mediate TUG localization to the ERGIC. a Schematic of TUG variants used for expression in TUG KO HeLa cells. All the proteins also contained a C-terminal mCherry tag to aid the visualization of the proteins in living cells. b Images of TUG KO HeLa cells transfected with full-length TUG (Tug-mCherry; left) or with a variant where the central IDR, IDR1 was deleted (Tug-IDR1∆-mCherry; right). Although full-length TUG forms punctate structures, the mutant is present in a diffuse distribution. Also note that the mutant is excluded from the nucleus. c Images of TUG KO HeLa cells transfected with IDR1 alone, tagged with mCherry (IDR1-mCherry; left), or with IDR2 alone, also tagged with mCherry (IDR2-mCherry; right). Although IDR1 forms punctate structures in cells, IDR2 is present in a diffuse distribution. In (b) and (c), a minimum of three independent transfections were performed for Tug-mCherry and IDR1-mCherry, three independent transfections for IDR2-mCherry and two independent transfections for Tug-IDR1∆-mCherry. d Images of TUG KO HeLa cells transfected with IDR1-mCherry and treated with 1,6-hexanediol and imaged before (left) or 150 sec. after 1,6-hexanediol treatment (right). Note that the punctate structures containing IDR1-mCherry continue to be present after 1,6-hexanediol treatment. The structures before and after are not necessarily at the same position, due to shift and refocusing during imaging after drug treatment. The treatment with 1,6-hexanediol was done three times, with similar results each time. e Images of TUG KO HeLa cells transfected with IDR1-mCherry (left) and mCherry-tagged IDR from FUS protein (right). Note the differences in the distribution of the two IDRs in cells. mCherry-tagged FUS IDR was transfected in TUG KO HeLa cells in two independent experiments, with similar results. f Images from TUG KO HeLa cells co-transfected with IDR1-mCherry (red) and Em-ERGIC53 (green). Note the colocalization of the two proteins in peripheral punctate structures. g Images from TUG KO HeLa cells co-transfected with IDR1-mCherry (red) and Stx12-GFP (green). Note the partial colocalization of the two proteins in punctate structures. Representative images for colocalization of IDR1-mCherry were from three independent experiments with Em-ERGIC53 (f) and two independent experiments with Stx12-GFP (g).CC BY · PubMed Central
Fig. 5 TUG oligomerizes in trans via its N-terminal ubiquitin-like domains. a Images of TUG KO MEFs infected with retroviruses to express mCherry-tagged TUG (magenta) tethered to mitochondria by the fusion of a transmembrane domain from the protein mitochondrial monoamine oxidase A (Tug-mCherry-Mito). Living cells were also incubated with Mitotracker green (green), which was washed off prior to imaging. Tethering of TUG to mitochondria results in mitochondrial clumping. b Images of TUG KO MEFs expressing mCherry tethered to mitochondria (magenta), together with Mitotracker green (green). Mitochondria were present in a filamentous organization under this condition. c, d Electron micrographs from TUG KO MEFs expressing Tug-mCherry-Mito (c) or without any exogenous protein expression (control; d) are shown. Mitochondria appear stacked when TUG is tethered to the mitochondrial outer membrane by monoamine oxidase A, but are dispersed in control cells. e–h Images of TUG KO MEFs expressing different truncations of TUG tagged with mCherry and tethered to the mitochondria. Deletion of IDR1 and IDR2 shows that these regions are not required for clumping of mitochondria (e); IDR1 alone is not sufficient to clump mitochondria (f). The N-terminal structured UBL1 and UBL2 regions in TUG protein are sufficient to mediate protein-protein interaction in trans and thus to clump mitochondria when fused to the transmembrane domain of mitochondrial monoamine oxidase A (g, h). All the above images are from TUG KO MEFs stably expressing the constructs; cells were imaged in two independent experiments. i, j Purified mCherry-tagged TUG (i) and a truncated version (deletion of N-terminal UBL1 and UBL2 domains, ( j) were incubated in the presence of different concentrations of Ficoll 400 and the condensates were imaged at same settings using confocal microscopy. Based on the size and intensity of the condensates, it is evident that the full-length protein is more effective in condensate formation compared to the deletion mutant, implying the structured UBL domains function to promote TUG condensation. Images with 5% Ficoll 400 were all acquired at similar setting for the wildtype and the mutant protein. Images with 2.5% Ficoll are acquired in the same imaging settings for the wildtype and the mutant protein. Experiments were repeated at least twice with similar results.
Fig. 5 TUG oligomerizes in trans via its N-terminal ubiquitin-like domains. a Images of TUG KO MEFs infected with retroviruses to express mCherry-tagged TUG (magenta) tethered to mitochondria by the fusion of a transmembrane domain from the protein mitochondrial monoamine oxidase A (Tug-mCherry-Mito). Living cells were also incubated with Mitotracker green (green), which was washed off prior to imaging. Tethering of TUG to mitochondria results in mitochondrial clumping. b Images of TUG KO MEFs expressing mCherry tethered to mitochondria (magenta), together with Mitotracker green (green). Mitochondria were present in a filamentous organization under this condition. c, d Electron micrographs from TUG KO MEFs expressing Tug-mCherry-Mito (c) or without any exogenous protein expression (control; d) are shown. Mitochondria appear stacked when TUG is tethered to the mitochondrial outer membrane by monoamine oxidase A, but are dispersed in control cells. e–h Images of TUG KO MEFs expressing different truncations of TUG tagged with mCherry and tethered to the mitochondria. Deletion of IDR1 and IDR2 shows that these regions are not required for clumping of mitochondria (e); IDR1 alone is not sufficient to clump mitochondria (f). The N-terminal structured UBL1 and UBL2 regions in TUG protein are sufficient to mediate protein-protein interaction in trans and thus to clump mitochondria when fused to the transmembrane domain of mitochondrial monoamine oxidase A (g, h). All the above images are from TUG KO MEFs stably expressing the constructs; cells were imaged in two independent experiments. i, j Purified mCherry-tagged TUG (i) and a truncated version (deletion of N-terminal UBL1 and UBL2 domains, ( j) were incubated in the presence of different concentrations of Ficoll 400 and the condensates were imaged at same settings using confocal microscopy. Based on the size and intensity of the condensates, it is evident that the full-length protein is more effective in condensate formation compared to the deletion mutant, implying the structured UBL domains function to promote TUG condensation. Images with 5% Ficoll 400 were all acquired at similar setting for the wildtype and the mutant protein. Images with 2.5% Ficoll are acquired in the same imaging settings for the wildtype and the mutant protein. Experiments were repeated at least twice with similar results.CC BY · PubMed Central
Fig. 6 Physiological effects of TUG deletion on autophagy, collagen secretion and CFTR trafficking. a In-gel fluorescence was used to monitor autophagic flux in WT and TUG KO MEFs. Processed and processed LC3 reporter bands were quantified. Note the low intensity of the processed band in KO MEFs. b, c Quantification of the ratio of processed to unprocessed LC3 bands (b) and ratio of intensities of processed to total LC3 reporter (c) from N = 4 independent experiments, normalized with respect to WT cells. Mean ± s.d., analyzed using a two-tailed t test. d Confocal images of WT and TUG KO MEFs, starved for 4 h in EBSS with BafA1 and stained for endogenous LC3B. Note the decreased LC3B in KO cells. e LC3B intensities per cell are plotted, normalized to the mean in WT cells. Medians are indicated. N = 70 WT and 61 KO cells from two independent experiments, analyzed using a two-tailed t test with Welch’s correction. f, g Abundances of secreted collagens (f) and control proteins (g) were quantified using mass spectrometry. Note the decreased collagen secretion in TUG KO MEFs. N = 4 independent experiments; mean ± s.d., analyzed using a two-tailed t test adjusted for multiple comparisons across all secreted proteins. h Images from WT and TUG KO MEFs transfected with GFP-tagged CFTR and stained using GFP booster (green) and an antibody to GM130. GFP-CFTR accumulates at the Golgi in TUG KO MEFs. i Pearson’s correlation coefficient was used to quantify the overlap of GFP-CFTR and GM130. N = 28 WT and 28 KO cells from three independent experiments. Mean ± s.d., analyzed by a two-tailed t test with Welch’s correction. Note that Pearson’s coefficient was increased in TUG KO cell, indicating that CFTR accumulates at the Golgi. j Immunoblot to detect abundances of ectopically expressed signal sequence-GFP (ssGFP) in cell lysates and secreted into media in WT and TUG KO MEFs. k Plots show the amount of secreted GFP, normalized to that in cell lysates. Data are normalized to WT cells. N = 3 independent experiments. Mean ± s.d., analyzed by a two-tailed t test with Welch’s correction.
Fig. 6 Physiological effects of TUG deletion on autophagy, collagen secretion and CFTR trafficking. a In-gel fluorescence was used to monitor autophagic flux in WT and TUG KO MEFs. Processed and processed LC3 reporter bands were quantified. Note the low intensity of the processed band in KO MEFs. b, c Quantification of the ratio of processed to unprocessed LC3 bands (b) and ratio of intensities of processed to total LC3 reporter (c) from N = 4 independent experiments, normalized with respect to WT cells. Mean ± s.d., analyzed using a two-tailed t test. d Confocal images of WT and TUG KO MEFs, starved for 4 h in EBSS with BafA1 and stained for endogenous LC3B. Note the decreased LC3B in KO cells. e LC3B intensities per cell are plotted, normalized to the mean in WT cells. Medians are indicated. N = 70 WT and 61 KO cells from two independent experiments, analyzed using a two-tailed t test with Welch’s correction. f, g Abundances of secreted collagens (f) and control proteins (g) were quantified using mass spectrometry. Note the decreased collagen secretion in TUG KO MEFs. N = 4 independent experiments; mean ± s.d., analyzed using a two-tailed t test adjusted for multiple comparisons across all secreted proteins. h Images from WT and TUG KO MEFs transfected with GFP-tagged CFTR and stained using GFP booster (green) and an antibody to GM130. GFP-CFTR accumulates at the Golgi in TUG KO MEFs. i Pearson’s correlation coefficient was used to quantify the overlap of GFP-CFTR and GM130. N = 28 WT and 28 KO cells from three independent experiments. Mean ± s.d., analyzed by a two-tailed t test with Welch’s correction. Note that Pearson’s coefficient was increased in TUG KO cell, indicating that CFTR accumulates at the Golgi. j Immunoblot to detect abundances of ectopically expressed signal sequence-GFP (ssGFP) in cell lysates and secreted into media in WT and TUG KO MEFs. k Plots show the amount of secreted GFP, normalized to that in cell lysates. Data are normalized to WT cells. N = 3 independent experiments. Mean ± s.d., analyzed by a two-tailed t test with Welch’s correction.CC BY · PubMed Central

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Show all 2 references

Research works printed but not linked: 88

  1. Barlowe, C. & Helenius, A. Cargo capture and bulk flow in the early secretory pathway. Annu. Rev. Cell Dev. Biol. 32, 197–222 (2016). · DOI
  2. Appenzeller-Herzog, C. & Hauri, H. P. The ER-Golgi intermediate compartment (ERGIC): in search of its identity and function. J. Cell Sci. 119, 2173–2183 (2006). · DOI
  3. Saraste, J. & Marie, M. Intermediate compartment (IC): from pre-Golgi vacuoles to a semi-autonomous membrane system. Histochem Cell Biol. 150, 407–430 (2018). · DOI
  4. Weigel, A. V. et al. ER-to-Golgi protein delivery through an interwoven, tubular network extending from ER. Cell 184, 2412–2429.e2416 (2021). · DOI
  5. Tojima, T., Suda, Y., Jin, N., Kurokawa, K. & Nakano, A. Spatiotemporal dissection of the Golgi apparatus and the ER-Golgi intermediate compartment in budding yeast. Elife 13 (2024). · DOI
  6. Han, Y., Li, S. & Ge, L. Biogenesis of autophagosomes from the ERGIC membrane system. J. Genet. Genomics 50, 3–6 (2023). · DOI
  7. Ge, L., Melville, D., Zhang, M. & Schekman, R. The ER-Golgi intermediate compartment is a key membrane source for the LC3 lipidation step of autophagosome biogenesis. Elife 2, e00947 (2013). · DOI
  8. Chieregatti, E. & Meldolesi, J. Regulated exocytosis: new organelles for non-secretory purposes. Nat. Rev. Mol. Cell Biol. 6, 181–187 (2005). · DOI
  9. Grieve, A. G. & Rabouille, C. Golgi bypass: skirting around the heart of classical secretion. Cold Spring Harb. Perspect. Biol. 3 (2011). · DOI
  10. Gee, H. Y., Noh, S. H., Tang, B. L., Kim, K. H. & Lee, M. G. Rescue of DeltaF508-CFTR trafficking via a GRASP-dependent unconventional secretion pathway. Cell 146, 746–760 (2011). · DOI
  11. Yoo, J. S. et al. Non-conventional trafficking of the cystic fibrosis transmembrane conductance regulator through the early secretory pathway. J. Biol. Chem. 277, 11401–11409 (2002). · DOI
  12. Bogan, J. S. Regulation of glucose transporter translocation in health and diabetes. Annu. Rev. Biochem. 81, 507–532 (2012). · DOI · Paper page
  13. Li, D. T. et al. GLUT4 Storage vesicles: specialized organelles for regulated trafficking. Yale J. Biol. Med. 92, 453–470 (2019).
  14. Gould, G. W., Brodsky, F. M. & Bryant, N. J. Building GLUT4 vesicles: CHC22 Clathrin’s human touch. Trends Cell Biol. 30, 705–719 (2020). · DOI
  15. Greig, J. et al. CHC22 clathrin recruitment to the early secretory pathway requires two-site interaction with SNX5 and p115. EMBO J. 43, 4298–4323 (2024). · DOI
  16. Bogan, J. S., Hendon, N., McKee, A. E., Tsao, T. S. & Lodish, H. F. Functional cloning of TUG as a regulator of GLUT4 glucose transporter trafficking. Nature 425, 727–733 (2003). · DOI
  17. Yu, C., Cresswell, J., Loffler, M. G. & Bogan, J. S. The glucose transporter 4-regulating protein TUG is essential for highly insulin-responsive glucose uptake in 3T3-L1 adipocytes. J. Biol. Chem. 282, 7710–7722 (2007). · DOI
  18. Orme, C. M. & Bogan, J. S. The ubiquitin regulatory X (UBX) domain-containing protein TUG regulates the p97 ATPase and resides at the endoplasmic reticulum-golgi intermediate compartment. J. Biol. Chem. 287, 6679–6692 (2012). · DOI
  19. Bogan, J. S. et al. Endoproteolytic cleavage of TUG protein regulates GLUT4 glucose transporter translocation. J. Biol. Chem. 287, 23932–23947 (2012). · DOI
  20. Belman, J. P. et al. Acetylation of TUG protein promotes the accumulation of GLUT4 glucose transporters in an insulin-responsive intracellular compartment. J. Biol. Chem. 290, 4447–4463 (2015). · DOI
  21. Habtemichael, E. N. et al. Usp25m protease regulates ubiquitin-like processing of TUG proteins to control GLUT4 glucose transporter translocation in adipocytes. J. Biol. Chem. 293, 10466–10486 (2018). · DOI
  22. Bogan, J. S. Ubiquitin-like processing of TUG proteins as a mechanism to regulate glucose uptake and energy metabolism in fat and muscle. Front. Endocrinol. (Lausanne) 13, 1019405 (2022). · DOI
  23. Habtemichael, E. N. et al. Insulin-stimulated endoproteolytic TUG cleavage links energy expenditure with glucose uptake. Nat. Metab. 3, 378–393 (2021). · DOI
  24. Camus, S. M. et al. CHC22 clathrin mediates traffic from early secretory compartments for human GLUT4 pathway biogenesis. J Cell Biol, 219 (2020). · DOI
  25. Xu, Y. et al. Dual-mode of insulin action controls GLUT4 vesicle exocytosis. J. Cell Biol. 193, 643–653 (2011). · DOI
  26. Ladanyi, M. et al. The der(17)t(X;17)(p11;q25) of human alveolar soft part sarcoma fuses the TFE3 transcription factor gene to ASPL, a novel gene at 17q25. Oncogene 20, 48–57 (2001). · DOI
  27. Yan, R., Chen, K., Wang, B. & Xu, K. SURF4-induced tubular ERGIC selectively expedites ER-to-Golgi transport. Dev. Cell 57, 512–525.e518 (2022). · DOI
  28. Larance, M. et al. Characterization of the role of the Rab GTPase-activating protein AS160 in insulin-regulated GLUT4 trafficking. J. Biol. Chem. 280, 37803–37813 (2005). · DOI
  29. Jedrychowski, M. P. et al. Proteomic analysis of GLUT4 storage vesicles reveals LRP1 to be an important vesicle component and target of insulin signaling. J. Biol. Chem. 285, 104–114 (2010). · DOI
  30. Song, X., Lichti, C. F., Townsend, R. R. & Mueckler, M. Single point mutations result in the miss-sorting of Glut4 to a novel membrane compartment associated with stress granule proteins. PLoS ONE 8, e68516 (2013). · DOI
  31. Fazakerley, D. J. et al. Proteomic analysis of GLUT4 storage vesicles reveals tumor suppressor candidate 5 (TUSC5) as a novel regulator of insulin action in adipocytes. J. Biol. Chem. 290, 23528–23542 (2015). · DOI
  32. Ray, A. et al. Regulated dynamic subcellular GLUT4 localization revealed by proximal proteome mapping in human muscle cells. J. Cell Sci. 136 (2023). · DOI
  33. Wakana, Y. et al. Kinesin-5/Eg5 is important for transport of CARTS from the trans-Golgi network to the cell surface. J. Cell Biol. 202, 241–250 (2013). · DOI
  34. Lujan, P. et al. Sorting of secretory proteins at the trans-Golgi network by human TGN46. Elife 12 (2024). · DOI
  35. Saxena, S. et al. Endoplasmic reticulum exit sites are segregated for secretion based on cargo size. Dev. Cell (2024). · DOI
  36. Rivera, V. M. et al. Regulation of protein secretion through controlled aggregation in the endoplasmic reticulum. Science 287, 826–830 (2000). · DOI
  37. Ernst, A. M. et al. S-Palmitoylation sorts membrane cargo for anterograde transport in the golgi. Dev. Cell 47, 479 (2018). · DOI
  38. Huang, F. et al. Ultra-high resolution 3d imaging of whole cells. Cell 166, 1028–1040 (2016). · DOI
  39. Xue, B., Dunbrack, R. L., Williams, R. W., Dunker, A. K. & Uversky, V. N. PONDR-FIT: a meta-predictor of intrinsically disordered amino acids. Biochim. Biophys. Acta 1804, 996–1010 (2010). · DOI
  40. Liang, Q. et al. MolPhase, an advanced prediction algorithm for protein phase separation. EMBO J. 43, 1898–1918 (2024). · DOI
  41. Park, D. et al. Cooperative function of synaptophysin and synapsin in the generation of synaptic vesicle-like clusters in non-neuronal cells. Nat. Commun. 12, 263 (2021). · DOI
  42. Kato, M. & McKnight, S. L. A solid-state conceptualization of information transfer from gene to message to protein. Annu Rev. Biochem 87, 351–390 (2018). · DOI
  43. Lin, Y. et al. Toxic PR poly-dipeptides encoded by the C9orf72 repeat expansion target LC domain polymers. Cell 167, 789–802.e712 (2016). · DOI
  44. Patel, A. et al. A liquid-to-solid phase transition of the ALS protein FUS accelerated by disease mutation. Cell 162, 1066–1077 (2015). · DOI
  45. De La Cruz, N. et al. Disorder-mediated interactions target proteins to specific condensates. Mol. Cell 84, 3497–3512 e3499 (2024). · DOI
  46. Lyons, H. et al. Functional partitioning of transcriptional regulators by patterned charge blocks. Cell 186, 327–345 e328 (2023). · DOI
  47. Wang, J. et al. A molecular grammar governing the driving forces for phase separation of prion-like RNA binding proteins. Cell 174, 688–699 e616 (2018). · DOI
  48. Wong, M. & Munro, S. Membrane trafficking. The specificity of vesicle traffic to the Golgi is encoded in the golgin coiled-coil proteins. Science 346, 1256898 (2014). · DOI
  49. Su, X. et al. Phase separation of signaling molecules promotes T cell receptor signal transduction. Science 352, 595–599 (2016). · DOI
  50. Yim, W. W., Yamamoto, H. & Mizushima, N. A pulse-chasable reporter processing assay for mammalian autophagic flux with HaloTag. Elife 11 (2022). · DOI
  51. Aridor, M. A tango for coats and membranes: New insights into ER-to-Golgi traffic. Cell Rep. 38, 110258 (2022). · DOI
  52. Raote, I. & Malhotra, V. Tunnels for protein export from the endoplasmic reticulum. Annu. Rev. Biochem. 90, 605–630 (2021). · DOI
  53. Santos, A. J., Raote, I., Scarpa, M., Brouwers, N. & Malhotra, V. TANGO1 recruits ERGIC membranes to the endoplasmic reticulum for procollagen export. Elife 4 (2015). · DOI
  54. Gee, H. Y., Kim, J. & Lee, M. G. Unconventional secretion of transmembrane proteins. Semin Cell Dev. Biol. 83, 59–66 (2018). · DOI
  55. Cheng, J. et al. A Golgi-associated PDZ domain protein modulates cystic fibrosis transmembrane regulator plasma membrane expression. J. Biol. Chem. 277, 3520–3529 (2002). · DOI
  56. Cheng, J., Cebotaru, V., Cebotaru, L. & Guggino, W. B. Syntaxin 6 and CAL mediate the degradation of the cystic fibrosis transmembrane conductance regulator. Mol. Biol. Cell 21, 1178–1187 (2010). · DOI
  57. Rebane, A. A. et al. Liquid-liquid phase separation of the Golgi matrix protein GM130. FEBS Lett. 594, 1132–1144 (2020). · DOI
  58. Ziltener, P., Rebane, A. A., Graham, M., Ernst, A. M. & Rothman, J. E. The golgin family exhibits a propensity to form condensates in living cells. FEBS Lett. 594, 3086–3094 (2020). · DOI
  59. Zhang, Y. & Seemann, J. RNA scaffolds the Golgi ribbon by forming condensates with GM130. Nat. Cell Biol. 26, 1139–1153 (2024). · DOI
  60. Su, M. et al. The golgi rim is a precise tetraplex of golgin proteins that can self-assemble into filamentous bands. bioRxiv https://doi.org/10.1101/2025.03.27.645134 (2025). · DOI
  61. Gallo, R., Rai, A. K., McIntyre, A. B. R., Meyer, K. & Pelkmans, L. DYRK3 enables secretory trafficking by maintaining the liquid-like state of ER exit sites. Dev. Cell 58, 1880–1897.e1811 (2023). · DOI
  62. Wegeng, W. R. et al. A hollow TFG condensate spatially compartmentalizes the early secretory pathway. bioRxiv (2024). · DOI · Paper page
  63. Hanna, M. G., Peotter, J. L., Frankel, E. B. & Audhya, A. Membrane transport at an organelle interface in the early secretory pathway: take your coat off and stay a while: evolution of the metazoan early secretory pathway. Bioessays 40, e1800004 (2018). · DOI
  64. Peotter, J. L. et al. TFG regulates secretory and endosomal sorting pathways in neurons to promote their activity and maintenance. Proc. Natl Acad. Sci. USA 119, e2210649119 (2022). · DOI
  65. Johnson, A. et al. TFG clusters COPII-coated transport carriers and promotes early secretory pathway organization. EMBO J. 34, 811–827 (2015). · DOI
  66. Loffler, M. G. et al. Enhanced fasting glucose turnover in mice with disrupted action of TUG protein in skeletal muscle. J. Biol. Chem. 288, 20135–20150 (2013). · DOI
  67. Habtemichael, E. N. et al. Coordinated regulation of vasopressin inactivation and glucose uptake by action of TUG protein in muscle. J. Biol. Chem. 290, 14454–14461 (2015). · DOI
  68. Semiz, S. et al. Conventional kinesin KIF5B mediates insulin-stimulated GLUT4 movements on microtubules. EMBO J. 22, 2387–2399 (2003). · DOI
  69. Wang, Y. et al. Trans-Golgi network syntaxin 10 functions distinctly from syntaxins 6 and 16. Mol. Membr. Biol. 22, 313–325 (2005). · DOI
  70. Nielsen, E. et al. Rabenosyn-5, a novel Rab5 effector, is complexed with hVPS45 and recruited to endosomes through a FYVE finger domain. J. Cell Biol. 151, 601–612 (2000). · DOI
  71. Koike, S. & Jahn, R. SNAREs define targeting specificity of trafficking vesicles by combinatorial interaction with tethering factors. Nat. Commun. 10, 1608 (2019). · DOI
  72. Bisio, H. et al. The ZIP code of vesicle trafficking in apicomplexa: SEC1/Munc18 and SNARE proteins. mBio 11, (2020). · DOI
  73. Milovanovic, D., Wu, Y., Bian, X. & De Camilli, P. A liquid phase of synapsin and lipid vesicles. Science 361, 604–607 (2018). · DOI
  74. Park, D. et al. Synaptic vesicle proteins and ATG9A self-organize in distinct vesicle phases within synapsin condensates. Nat. Commun. 14, 455 (2023). · DOI
  75. Park, D. et al. Overlapping role of synaptophysin and synaptogyrin family proteins in determining the small size of synaptic vesicles. Proc. Natl Acad. Sci. USA 121, e2409605121 (2024). · DOI
  76. Leung, A. K. L. Poly(ADP-ribose): a dynamic trigger for biomolecular condensate formation. Trends Cell Biol. 30, 370–383 (2020). · DOI
  77. Rhine, K. et al. Poly(ADP-ribose) drives condensation of FUS via a transient interaction. Mol. Cell 82, 969–985 e911 (2022). · DOI
  78. Hendriks, I. A., Larsen, S. C. & Nielsen, M. L. An advanced strategy for comprehensive profiling of ADP-ribosylation sites using mass spectrometry-based proteomics. Mol. Cell Proteom. 18, 1010–1026 (2019). · DOI
  79. Liu, X. et al. Generation of mammalian cells stably expressing multiple genes at predetermined levels. Anal. Biochem 280, 20–28 (2000). · DOI
  80. Parchure, A. et al. Liquid-liquid phase separation facilitates the biogenesis of secretory storage granules. J Cell Biol. 221 (2022). · DOI
  81. Amick, J., Roczniak-Ferguson, A. & Ferguson, S. M. C9orf72 binds SMCR8, localizes to lysosomes, and regulates mTORC1 signaling. Mol. Biol. Cell 27, 3040–3051 (2016). · DOI
  82. Devereaux, K. et al. Regulation of mammalian autophagy by class II and III PI 3-kinases through PI3P synthesis. PLoS ONE 8, e76405 (2013). · DOI · Paper page
  83. Mastronarde, D. N. Correction for non-perpendicularity of beam and tilt axis in tomographic reconstructions with the IMOD package. J. Microsc. 230, 212–217 (2008). · DOI
  84. Zhang, Y. et al. Nanoscale subcellular architecture revealed by multicolor three-dimensional salvaged fluorescence imaging. Nat. Methods 17, 225–231 (2020). · DOI
  85. Naviaux, R. K., Costanzi, E., Haas, M. & Verma, I. M. The pCL vector system: rapid production of helper-free, high-titer, recombinant retroviruses. J. Virol. 70, 5701–5705 (1996). · DOI
  86. Marin, Z. et al. PYMEVisualize: an open-source tool for exploring 3D super-resolution data. Nat. Methods 18, 582–584 (2021). · DOI
  87. Marin, Z., Fuentes, L. A., Bewersdorf, J. & Baddeley, D. Extracting nanoscale membrane morphology from single-molecule localizations. Biophys. J. 122, 3022–3030 (2023). · DOI
  88. Cignoni, P. et al. MeshLab: an open-source mesh processing tool. In: Sixth Eurographics Italian Chapter Conference (eds Scarano, V., De Chiara, R. & Erra, U.) 129–136 (The Eurographics Association, 2008).

Metadata

Authors
Parchure, A., Tejada, H., Xi, Z., Kim, Y., Su, M., Yan, Y., Julca-Zevallos, O., Alcázar-Román, A. R., Villemeur, M., Liu, X., Toomre, D., Raote, I., & Bogan, J. S.
Affiliations
  • Yale University (US)
  • Centre National de la Recherche Scientifique (FR)
Corresponding author
  • Parchure, Anup
  • Bogan, Jonathan S.
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2025-07-01
Volume
16
Issue
1
Pages
5518-5518
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-025-60691-8
PMC
PMC12218103
OpenAlex
W4411886980
Cited by
5
References
2
Citations by year
2026: 3 · 2025: 1
FWCI
1.99
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • HAL (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • U.S. Department of Health & Human Services | NIH | National Institute of Diabetes and Digestive and Kidney Diseases (R01 DK129466, P30 DK045735)
  • Blavatnik Family Foundation
  • U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (R01 GM151829, R01 GM134148)
  • Fondation pour la Recherche Médicale (AJE202210016216)
  • Agence Nationale de la Recherche (MatSec)
  • Fondation ARC pour la Recherche sur le Cancer
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Autophagy in Disease and Therapy
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Endoplasmic Reticulum Stress and Disease
Keywords
ASPSCR1, ERGIC, early secretory pathway, intrinsically disordered regions, biomolecular condensates, protein oligomerization, Golgi morphology, cis-Golgi, autophagy, collagen secretion, GLUT4 trafficking, membrane trafficking
MeSH terms
Animals, Autophagy, Collagen, Endoplasmic Reticulum, Golgi Apparatus, Humans, Cystic Fibrosis Transmembrane Conductance Regulator, Protein Transport, COP-Coated Vesicles, Mice, Secretory Pathway

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review

How to cite

Cite CoChat for the checks on this page, and the paper for its own findings.

This page
CoChat. (2026). Citation check: TUG protein acts through a disordered region to organize the early secretory pathway. https://app.cochat.ai/papers/10.1038/s41467-025-60691-8
The paper
Parchure, A., Tejada, H., Xi, Z., Kim, Y., Su, M., Yan, Y., Julca-Zevallos, O., Alcázar-Román, A. R., Villemeur, M., Liu, X., Toomre, D., Raote, I., & Bogan, J. S. (2025). TUG protein acts through a disordered region to organize the early secretory pathway. Nature Communications. https://doi.org/10.1038/s41467-025-60691-8