CHC22 clathrin mediates traffic from early secretory compartments for human GLUT4 pathway biogenesis

Camus, Stéphane M.; Camus, Marine; Figueras-Novoa, Carmen; Boncompain, Gaëlle; Sadacca, L. Amanda; Esk, Christopher; Bigot, Anne; Gould, Gwyn W.; Kioumourtzoglou, Dimitrios; Perez, Franck; Bryant, Nia J.; Mukherjee, Shaeri; Brodsky, Frances M.

doi:10.1101/242941

Cited by 12 publications

(55 citation statements)

References 84 publications

(160 reference statements)

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“…indicates that CHC22 enhances membrane traffic in an existing transport route for GLUT4 from the secretory pathway such that this route becomes more efficient for delivering GLUT4 to its insulin-responsive storage compartment and complements the established endocytic (30). Thus, we suggest that species with functional CHC22 clathrin are better at intracellular GLUT4 sequestration, resulting in lower surface GLUT4 in the absence of insulin, and tighter regulation of GLUT4 release in response to insulin.…”

Section: Discussionmentioning

confidence: 85%

“…Atypically for their epithelial cell origin but not for transformed cells, HeLa cells express CHC22 clathrin (they are homozygous for the M1316 variant) (28,29). We observed that the transfected fluorescently tagged CHC22 variants both localized similarly to endogenous CHC22-Met1316 (30). The dynamics of membrane association for the two allotypes was then tested.…”

Section: Chc22 Variants Display Functional Differencesmentioning

confidence: 99%

“…These cells can sequester GLUT4 intracellularly, and then release it to the plasma membrane in response to insulin, behaving like muscle and adipocytes, though with more modest response (30)(31)(32). Transfection of HeLa cells with siRNA depleting CHC22 ablates this insulin-responsive pathway (30). To detect GLUT4 release to the cell surface, we used a construct expressing GLUT4 tagged with mCherry and a hemagglutinin (HA) epitope embedded in an exofacial loop of the transporter (HA-GLUT4-mCherry).…”

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confidence: 99%

“…Because HeLa cells express CHC22, they can form a GLUT4 storage compartment (GSC), when transfected to express GLUT4. These cells can sequester GLUT4 intracellularly, and then release it to the plasma membrane in response to insulin, behaving like muscle and adipocytes, though with more modest response (30)(31)(32). Transfection of HeLa cells with siRNA depleting CHC22 ablates this insulin-responsive pathway (30).…”

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confidence: 99%

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Genetic diversity of CHC22 clathrin impacts its function in glucose metabolism

Fumagalli

Camus

Diekmann

et al. 2018

Preprint

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CHC22 clathrin plays a key role in intracellular membrane trafficking of the insulinresponsive GLUT4 glucose transporter, and so in post-prandial clearance of glucose from human blood. We performed population genetic and phylogenetic analyses of the CLTCL1 gene, encoding CHC22, to understand its variable presence in vertebrates and gain insight into its functional evolution. Analysis of ~50 complete vertebrate genomes showed independent loss of CLTCL1 in nine lineages after it arose from a gene duplication during the emergence of jawed vertebrates. All vertebrates considered here retained the parent CLTC gene encoding CHC17 clathrin, which mediates endocytosis and other housekeeping membrane traffic pathways, as performed by the single clathrin gene product in nonvertebrate eukaryotes. Statistical analysis provides evidence of strong purifying selection over phylogenetic timescales for CLTCL1, as well as for CLTC, supporting preserved functionality of CHC22 in those species that have retained CLTCL1. In population genetic analyses of humans and chimpanzees, extensive allelic diversity was observed for CLTCL1 compared to CLTC. In all human populations, two variants of CLTCL1 segregate at high frequency, resulting in CHC22 protein with either methionine or valine at position 1316. The V1316 variant occurs only in humans, but the same site is polymorphic in non-human primates as well. Analysis of archaic and ancient humans assigned the appearance of the derived V1316 allele to 500-50 KYA. Balancing selection on the two high-frequency CHC22 variants is inferred, with V1316 being more frequent in farming, as compared to huntergatherer populations. Together these analyses suggest that CHC22 clathrin is undergoing selection in humans related to its role in nutrient metabolism. Consistent with this conclusion, we observed functional differences between the two CHC22 variants in their ability to control GLUT4 membrane traffic, as predicted by structural modeling and differences in cellular dynamics of the two variants.

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Section: Discussionmentioning

confidence: 85%

Section: Chc22 Variants Display Functional Differencesmentioning

confidence: 99%

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confidence: 99%

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Genetic diversity of CHC22 clathrin impacts its function in glucose metabolism

Fumagalli

Camus

Diekmann

et al. 2018

Preprint

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“…However, the enriched immunoabsorption of cargo whose trafficking is specialized, but not constitutively recycling cargo, argues we are able to distinguish different regions or subdomains of the TGN. Interestingly, when GLUT4 is ectopically expressed in cell types that do not natively express GLUT4, such as fibroblasts, CHO cells, and HeLa cells, an insulin regulated recycling mechanism does exist, albeit less robust than in adipocytes (12, 60). Specifically, it has been demonstrated that GLUT4 travels to the PM in vesicles that are distinct from vesicles carrying constitutively recycling cargo (12).…”

Section: Discussionmentioning

confidence: 99%

Insulin promoted mobilization of GLUT4 from a perinuclear storage site requires RAB10

Brumfield

Chaudhary

Molle

et al. 2020

Preprint

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25Insulin controls glucose uptake into muscle and fat cells by inducing a net redistribution of 26 GLUT4 from intracellular storage to the plasma membrane (PM). The TBC1D4-RAB10 signaling 27 module is required for insulin-stimulated GLUT4 translocation to the PM, although where it 28 intersects GLUT4 traffic was unknown. Here we demonstrate that TBC1D4-RAB10 functions to 29 control GLUT4 mobilization from a Trans Golgi Network (TGN) storage compartment, 30 establishing that insulin, in addition to regulating the PM proximal effects of GLUT4-containing 31 vesicles docking to and fusion with the PM, also directly regulates the behavior of GLUT4 32 deeper within the cell. We also show that GLUT4 is retained in an element/domain of the TGN 33 from which newly synthesized lysosomal proteins are targeted to the late endosomes and the 34 ATP7A copper transporter is translocated to the PM by elevated copper. Insulin does not 35 mobilize ATP7A nor does copper mobilize GLUT4. Consequently, GLUT4 intracellular 36 sequestration and mobilization by insulin is achieved, in part, through utilizing a region of the 37 TGN devoted to specialized cargo transport in general rather than being specific for GLUT4. 38Our results define GLUT4-containing region of the TGN as a sorting and storage site from which 39 different cargo are mobilized by distinct signals. 40 11) whose delivery to the PM is regulated by insulin (2). GLUT4 in the PM cycles back to the 51 TGN via the endosomal pathway (2, 12, 13). Targeting GLUT4 from endosomes to the TGN has 52 an important role in basal intracellular GLUT4 retention. Mutations in GLUT4 that disrupt its 53 traffic from endosomes to the TGN are poorly retained in basal conditions and are not properly 54 translocated to the PM upon insulin stimulation (6, 14-16). These results identify the TGN as the 55 site for formation of IRVs. The TGN is a main sorting compartment along the biosynthetic and 56 endocytic pathways. Cargoes to be targeted to distinct destinations are sorted and packaged 57 into the correct transport vesicles in the TGN. The relationship between the TGN containing 58 GLUT4 and the TGN involved in the traffic of other cargoes is not known (2, 6, 8, 12). 59Insulin signaling triggers multiple discrete molecular events that mediate efficient recruitment, 60 docking, and fusion of IRVs with the PM (17-19). These events lead to a decrease in the size of 61 the intracellular GLUT4 pool concomitant with an increase of GLUT4 in the PM. As GLUT4 in 62 the PM is in equilibrium with intracellular GLUT4, endocytosis of GLUT4 dynamically removes 63 GLUT4 from the PM. Thus, maintenance of the insulin-stimulated dynamic increase in GLUT4 64 in the PM requires the continual ferrying of GLUT4-containing IRVs to the PM. Insulin signaling 65 can add to the IRV pool by increasing the rate of GLUT4 mobilization from the TGN in nascent 66IRVs. Despite the biological importance, insulin regulation of GLUT4 trafficking at the 67 perinuclear region has not been thoroughly interrogated. 68A key aspect of insu...

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Diabetes is accompanied by changes in the levels of proteins involved in endosomal GLUT4 trafficking in obese human skeletal muscle

Livingstone

Bryant

Boyle

et al. 2022

Endocrino Diabet & Metabol

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Introduction The regulated delivery of the glucose transporter GLUT4 from intracellular stores to the plasma membrane underpins insulin‐stimulated glucose transport. Insulin‐stimulated glucose transport is impaired in skeletal muscle of patients with type‐2 diabetes, and this may arise because of impaired intracellular trafficking of GLUT4. However, molecular details of any such impairment have not been described. We hypothesized that GLUT4 and/or levels of proteins involved in intracellular GLUT4 trafficking may be impaired in skeletal muscle in type‐2 diabetes and tested this in obese individuals without and without type‐2 diabetes. Methods We recruited 12 participants with type‐2 diabetes and 12 control participants. All were overweight or obese with BMI of 25–45 kg/m2. Insulin sensitivity was measured using an insulin suppression test (IST), and vastus lateralis biopsies were taken in the fasted state. Cell extracts were immunoblotted to quantify levels of a range of proteins known to be involved in intracellular GLUT4 trafficking. Results Obese participants with type‐2 diabetes exhibited elevated fasting blood glucose and increased steady state glucose infusion rates in the IST compared with controls. Consistent with this, skeletal muscle from those with type‐2 diabetes expressed lower levels of GLUT4 (30%, p = .014). Levels of Syntaxin4, a key protein involved in GLUT4 vesicle fusion with the plasma membrane, were similar between groups. By contrast, we observed reductions in levels of Syntaxin16 (33.7%, p = 0.05), Sortilin (44%, p = .006) and Sorting Nexin‐1 (21.5%, p = .039) and −27 (60%, p = .001), key proteins involved in the intracellular sorting of GLUT4, in participants with type‐2 diabetes. Conclusions We report significant reductions of proteins involved in the endosomal trafficking of GLUT4 in skeletal muscle in obese people with type 2 diabetes compared with age‐ and weight‐matched controls. These abnormalities of intracellular GLUT4 trafficking may contribute to reduced whole body insulin sensitivity.

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CHC22 clathrin mediates traffic from early secretory compartments for human GLUT4 pathway biogenesis

Cited by 12 publications

References 84 publications

Genetic diversity of CHC22 clathrin impacts its function in glucose metabolism

Genetic diversity of CHC22 clathrin impacts its function in glucose metabolism

Insulin promoted mobilization of GLUT4 from a perinuclear storage site requires RAB10

Diabetes is accompanied by changes in the levels of proteins involved in endosomal GLUT4 trafficking in obese human skeletal muscle

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