Recycling of epidermal growth factor-receptor complexes in A431 cells: identification of dual pathways.

Sorkin, Alexander; Krolenko, Sergey; Kudrjavtceva, Natalia; Lazebnik, Juri; Teslenko, L V; Soderquist, Ann Mangelsdorf; Nikolsky, Nikolay

doi:10.1083/jcb.112.1.55

Cited by 96 publications

(70 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The present results demonstrating that chloroquine effectively interrupts the degradative processing without exerting a deleterious effect on the retroendocytotic pathway is novel and intriguing. In agreement with our findings, several other types of ligand-receptor complexes recycle through the chloroquine-insensitive pathway, including EGF, insulin, and asialoglycoprotein receptors (46,47,56,63,64). This establishes the notion that after internalization, many types of ligand-receptor complexes can recycle through the chloroquine-insensitive pathway and finally be degraded via chloroquine-sensitive lysosomal pathway.…”

Section: Discussionsupporting

confidence: 79%

Ligand-regulated Internalization, Trafficking, and Down-regulation of Guanylyl Cyclase/Atrial Natriuretic Peptide Receptor-A in Human Embryonic Kidney 293 Cells

Pandey

Nguyen

Sharma

et al. 2002

Journal of Biological Chemistry

128

View full text Add to dashboard Cite

We examined the kinetics of internalization, trafficking, and down-regulation of recombinant guanylyl cyclase/natriuretic peptide receptor-A (NPRA) utilizing stably transfected 293 cells expressing a very high density of receptors. After atrial natriuretic peptide (ANP) binding to NPRA, ligand-receptor complexes are internalized, processed intracellularly, and sequestered into subcellular compartments, which provided an approach to examining directly the dynamics of metabolic turnover of NPRA in intact cells. The translocation of ligandreceptor complexes from cell surface to intracellular compartments seems to be linked to ANP-dependent down-regulation of NPRA. Using tryptic proteolysis of cell surface receptors, it was found that ϳ40 -50% of internalized ligand-receptor complexes recycled back to the plasma membrane with an apparent t1 ⁄2 ‫؍‬ 8 min. The recycling of NPRA was blocked by the lysosomotropic agent chloroquine, the energy depleter dinitrophenol, and also by low temperature, suggesting that recycling of the receptor is an energy-and temperature-dependent process. Data suggest that ϳ70 -80% of internalized 125 I-ANP is processed through a lysosomal degradative pathway; however, 20 -25% of internalized ligand is released intact into the cell exterior through an alternative mechanism involving an chloroquine-insensitive pathway. It is implied that internalization and processing of bound ANP-NPRA complexes may play an important role in mediating the biological action of hormone and the receptor protein. In retrospect, this could occur at the level of receptor regulation or through the initiation of ANP mediated signals. It is envisioned that the endocytotic pathway of ligand-receptor complexes of ANP-NPRA would lead to termination and/or diminished responsiveness of ANP in target cells.

show abstract

Section: Discussionsupporting

confidence: 79%

Ligand-regulated Internalization, Trafficking, and Down-regulation of Guanylyl Cyclase/Atrial Natriuretic Peptide Receptor-A in Human Embryonic Kidney 293 Cells

Pandey

Nguyen

Sharma

et al. 2002

Journal of Biological Chemistry

128

View full text Add to dashboard Cite

show abstract

“…S2A-C and S3), which blocks trafficking towards degradation compartments and impairs recycling towards the plasma membrane (Futter et al, 1996;Sorkin et al, 1991a). The amount of EGF was higher in TI-VAMP-depleted cells with both TI-VAMP siRNAs and not with the silencing of other v-SNAREs (VAMP3, VAMP4 or VAMP8; supplementary material Figs S2C, S4).…”

Section: Ti-vamp Depletion Accelerates Endocytosis Of Activated Egfrmentioning

confidence: 91%

Role of TI-VAMP and CD82 in EGFR cell-surface dynamics and signaling

Danglot

Chaineau

Dahan

et al. 2010

Journal of Cell Science

View full text Add to dashboard Cite

SummaryThe v-SNARE TI-VAMP (VAMP7) mediates exocytosis during neuritogenesis, phagocytosis and lysosomal secretion. It localizes to endosomes and lysosomes but also to the trans-Golgi network. Here we show that depletion of TI-VAMP enhances the endocytosis of activated EGF receptor (EGFR) without affecting constitutive endocytosis of EGFR, or transferrin uptake. This increased EGFR internalization is mainly clathrin dependent. Searching for defects in EGFR regulators, we found that TI-VAMP depletion reduces the cell surface amount of CD82, a tetraspanin known to control EGFR localization in microdomains. We further show that TI-VAMP is required for secretion from the Golgi apparatus to the cell surface, and that TI-VAMP-positive vesicles transport CD82. Quantum dots video-microscopy indicates that depletion of TI-VAMP, or its cargo CD82, restrains EGFR diffusion and the area explored by EGFR at the cell surface. Both depletions also impair MAPK signaling and enhance endocytosis of activated EGFR by increased recruitment of AP-2. These results highlight the role of TI-VAMP in the secretory pathway of a tetraspanin, and support a model in which CD82 allows EGFR entry in microdomains that control its clathrin-dependent endocytosis and signaling.Key words: CD82, EGFR, Endocytosis, Tetraspanin, TI-VAMP, L1-CAM, AP-2 Journal of Cell Science 724microdomains that control its clathrin-dependent endocytosis and regulate its signaling. Results TI-VAMP depletion accelerates endocytosis of activated EGFRTo determine whether TI-VAMP knockdown (KD) had an effect on the endocytic pathway we evaluated EGFR endocytosis kinetics by surface immunochemistry and flow cytometry. We silenced the expression of TI-VAMP in HeLa cells by either targeting the coding region (99% of extinction) or the 3Ј-UTR of TI-VAMP mRNA (97% of extinction, supplementary material Fig. S1). Efficient silencing required 96 hours incubation and was effective for at least 8 days.Surface EGFR was labeled at 4°C with an antibody which does not interfere with the binding of EGF. In the absence of EGF, the amount of EGFR internalized was not significantly different in mock-and TI-VAMP-depleted cells even after 30 minutes of endocytosis (Fig. 1A, left), indicating that constitutive endocytosis of EGFR was unchanged. In the presence of EGF, however, EGFR endocytosis was significantly enhanced in TI-VAMP-depleted cells (Fig. 1A, right). These results indicate that TI-VAMP depletion accelerates endocytosis of activated EGFR within a few minutes.The increased endocytosis was not due to an effect on expression of EGFR, since no significant difference in either the total level of EGFR, as determined by western blot (supplementary material Fig. S2E) or in the surface level of EGFR, quantified by cytometry (supplementary material Fig. S2F) was observed. Moreover we did not find any effect of TI-VAMP KD on transferrin internalization, indicating that TI-VAMP KD does not have a general stimulatory effect on cell endocytosis (Fig. 1B).The increased EGFR endocytosis was conf...

show abstract

“…These results also suggest that spartin might be involved in the internalization of EGFR. Alternatively, spartin depletion might instead influence the rate of EGFR recycling, which is known to have a half-time of ϳ5 min (Sorkin et al, 1991). Future detailed biochemical assays as well as experiments examining the presence of spartin on coated-pits will clarify the role of spartin in the internalization and/or recycling of EGFR.…”

Section: Discussionmentioning

confidence: 99%

Troyer Syndrome Protein Spartin Is Mono-Ubiquitinated and Functions in EGF Receptor Trafficking

Bakowska

Jupille

Fatheddin

et al. 2007

MBoC

133

View full text Add to dashboard Cite

Troyer syndrome is an autosomal recessive hereditary spastic paraplegia caused by mutation in the spartin (SPG20) gene, which encodes a widely expressed protein of unknown function. This mutation results in premature protein truncation and thus might signify a loss-of-function disease mechanism. In this study, we have found that spartin is monoubiquitinated and functions in degradation of the epidermal growth factor receptor (EGFR). Upon EGF stimulation, spartin translocates from the cytoplasm to the plasma membrane and colocalizes with internalized EGF-Alexa. Knockdown of spartin by small interfering RNA decreases the rate of EGFR degradation and also affects EGFR internalization, recycling, or both. Furthermore, overexpression of spartin results in a prominent decrease in EGFR degradation. Taken together, our data suggest that spartin is involved in the intracellular trafficking of EGFR and that impaired endocytosis may underlie the pathogenesis of Troyer syndrome. INTRODUCTIONThe hereditary spastic paraplegias (HSPs) comprise a cluster of inherited neurological disorders characterized by progressive spasticity and muscle weakness in the lower limbs Reid, 2003;Fink, 2006;Soderblom and Blackstone, 2006). Classically, the HSPs have been divided into two forms: "pure" when lower extremity spasticity and paraparesis are the only features and "complicated" when additional symptoms are present (Harding, 1983). More than 30 genetic loci (SPG1-33) and 14 gene products have been identified, yielding new insights into the molecular pathways involved in the pathogenesis of the HSPs. Troyer syndrome (SPG20), an autosomal recessive, complicated HSP that manifests in early childhood, is characterized by dysarthria, mental retardation, shortness of stature, and distal muscle wasting in addition to spasticity and weakness of the lower limbs (Cross and McKusick, 1967;Proukakis et al., 2004). To date, the only known mutation involved in Troyer syndrome is a single base deletion in the spartin gene, resulting in a 29-amino acid substitution at the C-terminus and premature truncation of the 666-amino acid protein by 268 residues (Patel et al., 2002;Proukakis et al., 2004).Although the cellular functions of spartin are not known, it harbors two conserved domains, an MIT (contained within microtubule-interacting and trafficking molecules) domain at the N-terminus and a plant-related region at the C-terminus. Although the only known function of the latter domain is in senescence (Panavas et al., 1999), the MIT domain is found in several other proteins, including Vps4-A and -B, sortin 15, and spastin (Ciccarelli et al., 2003), which are involved in vesicle trafficking and binding of microtubules. Interestingly, spastin, a microtubule-severing ATPase (Errico et al., 2002;Trotta et al., 2004;Evans et al., 2005), is mutated in the most common form of autosomal dominant HSP, SPG4 Reid, 2003;Fink, 2006;Soderblom and Blackstone, 2006). Although a region adjacent to the MIT domain is involved in the interaction of spastin with microtubules ...

show abstract

Recycling of epidermal growth factor-receptor complexes in A431 cells: identification of dual pathways.

Cited by 96 publications

References 42 publications

Ligand-regulated Internalization, Trafficking, and Down-regulation of Guanylyl Cyclase/Atrial Natriuretic Peptide Receptor-A in Human Embryonic Kidney 293 Cells

Ligand-regulated Internalization, Trafficking, and Down-regulation of Guanylyl Cyclase/Atrial Natriuretic Peptide Receptor-A in Human Embryonic Kidney 293 Cells

Role of TI-VAMP and CD82 in EGFR cell-surface dynamics and signaling

Troyer Syndrome Protein Spartin Is Mono-Ubiquitinated and Functions in EGF Receptor Trafficking

Contact Info

Product

Resources

About