Wortmannin-sensitive trafficking pathways in Chinese hamster ovary cells. Differential effects on endocytosis and lysosomal sorting - PubMed (original) (raw)
. 1996 May 3;271(18):10953-62.
doi: 10.1074/jbc.271.18.10953.
Affiliations
- PMID: 8631914
- DOI: 10.1074/jbc.271.18.10953
Free article
Wortmannin-sensitive trafficking pathways in Chinese hamster ovary cells. Differential effects on endocytosis and lysosomal sorting
J L Martys et al. J Biol Chem. 1996.
Free article
Abstract
Phosphatidylinositol (PI) 3'-kinases are a family of lipid kinases implicated in the regulation of cell growth by oncogene products and tyrosine kinase growth factor receptors. The catalytic subunit of the p85/p110 PI 3'-kinase is homologous to VPS-34, a phosphatidylinositol-specific lipid kinase involved in the sorting of newly synthesized hydrolases to the yeast vacuole. This suggests that PI 3'-kinases may play analogous roles in mammalian cells. We have measured a number of secretory and endocytic trafficking events in Chinese hamster ovary cells in the presence of wortmannin, a potent inhibitor of PI 3'-kinase. Wortmannin caused a 40-50% down-regulation of surface transferrin receptors, with a dose dependence identical to that required for maximal inhibition of the p85/p110 PI 3'-kinase in intact cells. The redistribution of transferrin receptors reflected a 60% increase in the internalization rate and a 35% decrease in the recycling rate. Experiments with fluorescent transferrin showed that entry of transferrin receptors into the recycling compartment and efflux of receptors out of the compartment were slowed by wortmannin. Wortmannin altered the morphology of the recycling compartment, which was more vesiculated than in untreated cells. Using Semliki Forest virus as a probe, we also found that delivery of the endocytosed virus to its lysosomal site of degradation was slowed by wortmannin, whereas endosomal acidification was unaffected. In contrast to these effects on endocytosis and recycling, wortmannin did not affect intracellular processing of newly synthesized viral spike proteins. Wortmannin did induce missorting of the lysosomal enzyme cathepsin D to the secretory pathway, but only at a dose 20-fold greater than that required to inhibit p85/p110 PI 3'-kinase activity or to redistribute transferrin receptors. Our data demonstrate the presence of wortmannin-sensitive enzymes at three distinct steps of the endocytic cycle in Chinese hamster ovary cells: internalization, transit from early endosomes to the recycling and degradative compartments, and transit from the recycling compartment back to the cell surface. The wortmannin-sensitive enzymes critical for endocytosis and recycling are distinct from those involved in sorting newly synthesized lysosomal enzymes.
Similar articles
- Potential sites of PI-3 kinase function in the endocytic pathway revealed by the PI-3 kinase inhibitor, wortmannin.
Shpetner H, Joly M, Hartley D, Corvera S. Shpetner H, et al. J Cell Biol. 1996 Feb;132(4):595-605. doi: 10.1083/jcb.132.4.595. J Cell Biol. 1996. PMID: 8647891 Free PMC article. - Role for phosphatidylinositol 3-kinase in the sorting and transport of newly synthesized lysosomal enzymes in mammalian cells.
Brown WJ, DeWald DB, Emr SD, Plutner H, Balch WE. Brown WJ, et al. J Cell Biol. 1995 Aug;130(4):781-96. doi: 10.1083/jcb.130.4.781. J Cell Biol. 1995. PMID: 7642697 Free PMC article. - The role of calcium and other ions in sorting and delivery in the late endocytic pathway.
Luzio JP, Bright NA, Pryor PR. Luzio JP, et al. Biochem Soc Trans. 2007 Nov;35(Pt 5):1088-91. doi: 10.1042/BST0351088. Biochem Soc Trans. 2007. PMID: 17956286 Review. - Small molecules targeting endocytic uptake and recycling pathways.
Placidi G, Mattu C, Ciardelli G, Campa CC. Placidi G, et al. Front Cell Dev Biol. 2023 Mar 10;11:1125801. doi: 10.3389/fcell.2023.1125801. eCollection 2023. Front Cell Dev Biol. 2023. PMID: 36968200 Free PMC article. Review.
Cited by
- Visualizing molecules of functional human profilin.
Pimm ML, Liu X, Tuli F, Heritz J, Lojko A, Henty-Ridilla JL. Pimm ML, et al. Elife. 2022 Jun 6;11:e76485. doi: 10.7554/eLife.76485. Elife. 2022. PMID: 35666129 Free PMC article. - Crossing the Iron Gate: Why and How Transferrin Receptors Mediate Viral Entry.
Wessling-Resnick M. Wessling-Resnick M. Annu Rev Nutr. 2018 Aug 21;38:431-458. doi: 10.1146/annurev-nutr-082117-051749. Epub 2018 May 31. Annu Rev Nutr. 2018. PMID: 29852086 Free PMC article. Review. - E3 Ubiquitin Ligase RNF125 Activates Interleukin-36 Receptor Signaling and Contributes to Its Turnover.
Saha SS, Caviness G, Yi G, Raymond EL, Mbow ML, Kao CC. Saha SS, et al. J Innate Immun. 2018;10(1):56-69. doi: 10.1159/000481210. Epub 2017 Nov 25. J Innate Immun. 2018. PMID: 29176319 Free PMC article. - Transferrin: Endocytosis and Cell Signaling in Parasitic Protozoa.
Reyes-López M, Piña-Vázquez C, Serrano-Luna J. Reyes-López M, et al. Biomed Res Int. 2015;2015:641392. doi: 10.1155/2015/641392. Epub 2015 May 18. Biomed Res Int. 2015. PMID: 26090431 Free PMC article. Review. - Profilin as a regulator of the membrane-actin cytoskeleton interface in plant cells.
Sun T, Li S, Ren H. Sun T, et al. Front Plant Sci. 2013 Dec 19;4:512. doi: 10.3389/fpls.2013.00512. Front Plant Sci. 2013. PMID: 24391654 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Research Materials
Miscellaneous