PtdIns(4,5)P2 Turnover is Required for Multiple Stages During Clathrin- and Actin-dependent Endocytic Internalization

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 2007 Apr 25

PMID 17452534

Citations 87

Authors

Yidi Sun

Susheela Carroll

Marko Kaksonen

Junko Y Toshima

David G Drubin

Affiliations

Soon will be listed here.

Abstract

The lipid phosphatidylinositol-4,5-bisphosphate (PtdIns[4,5]P(2)) appears to play an important role in endocytosis. However, the timing of its formation and turnover, and its specific functions at different stages during endocytic internalization, have not been established. In this study, Sla2 ANTH-GFP and Sjl2-3GFP were expressed as functional fusion proteins at endogenous levels to quantitatively explore PtdIns(4,5)P(2) dynamics during endocytosis in yeast. Our results indicate that PtdIns(4,5)P(2) levels increase and decline in conjunction with coat and actin assembly and disassembly, respectively. Live-cell image analysis of endocytic protein dynamics in an sjl1Delta sjl2Delta mutant, which has elevated PtdIns(4,5)P(2) levels, revealed that the endocytic machinery is still able to assemble and disassemble dynamically, albeit nonproductively. The defects in the dynamic behavior of the various endocytic proteins in this double mutant suggest that PtdIns(4,5)P(2) turnover is required for multiple stages during endocytic vesicle formation. Furthermore, our results indicate that PtdIns(4,5)P(2) turnover may act in coordination with the Ark1/Prk1 protein kinases in stimulating disassembly of the endocytic machinery.

Citing Articles

Phosphoinositide Depletion and Compensatory β-adrenergic Signaling in Angiotensin II-Induced Heart Disease: Protection Through PTEN Inhibition.

Voelker T, Voelker T, Westhoff M, Del Villar S, Thai P, Chiamvimonvat N bioRxiv. 2025; .

PMID: 40060428 PMC: 11888262. DOI: 10.1101/2025.02.23.639781.

The conserved protein adaptors CALM/AP180 and FCHo1/2 cooperatively recruit Eps15 to promote the initiation of clathrin-mediated endocytosis in yeast.

Sun Y, Yeam A, Kuo J, Iwamoto Y, Hu G, Drubin D PLoS Biol. 2024; 22(9):e3002833.

PMID: 39316607 PMC: 11451990. DOI: 10.1371/journal.pbio.3002833.

Endolysosomal trafficking controls yolk granule biogenesis in vitellogenic Drosophila oocytes.

Yu Y, Chen D, Farmer S, Xu S, Rios B, Solbach A PLoS Genet. 2024; 20(2):e1011152.

PMID: 38315726 PMC: 10898735. DOI: 10.1371/journal.pgen.1011152.

Mechanism of actin capping protein recruitment and turnover during clathrin-mediated endocytosis.

Lamb A, Fernandez A, Eadaim A, Johnson K, Di Pietro S J Cell Biol. 2023; 223(1).

PMID: 37966720 PMC: 10651396. DOI: 10.1083/jcb.202306154.

PLC regulates spontaneous glutamate release triggered by extracellular calcium and readily releasable pool size in neocortical neurons.

Feldthouse M, Vyleta N, Smith S Front Cell Neurosci. 2023; 17:1193485.

PMID: 37260580 PMC: 10228687. DOI: 10.3389/fncel.2023.1193485.

References

Cremona O, Di Paolo G, Wenk M, Luthi A, Kim W, Takei K . Essential role of phosphoinositide metabolism in synaptic vesicle recycling. Cell. 1999; 99(2):179-88. DOI: 10.1016/s0092-8674(00)81649-9. View

Stefan C, Audhya A, Emr S . The yeast synaptojanin-like proteins control the cellular distribution of phosphatidylinositol (4,5)-bisphosphate. Mol Biol Cell. 2002; 13(2):542-57. PMC: 65648. DOI: 10.1091/mbc.01-10-0476. View

HARRIS T, Hartwieg E, Horvitz H, Jorgensen E . Mutations in synaptojanin disrupt synaptic vesicle recycling. J Cell Biol. 2000; 150(3):589-600. PMC: 2175188. DOI: 10.1083/jcb.150.3.589. View

Gad H, Ringstad N, Low P, Kjaerulff O, Gustafsson J, Wenk M . Fission and uncoating of synaptic clathrin-coated vesicles are perturbed by disruption of interactions with the SH3 domain of endophilin. Neuron. 2000; 27(2):301-12. DOI: 10.1016/s0896-6273(00)00038-6. View

Itoh T, Koshiba S, Kigawa T, Kikuchi A, Yokoyama S, Takenawa T . Role of the ENTH domain in phosphatidylinositol-4,5-bisphosphate binding and endocytosis. Science. 2001; 291(5506):1047-51. DOI: 10.1126/science.291.5506.1047. View

Howard J, Hutton J, Olson J, Payne G . Sla1p serves as the targeting signal recognition factor for NPFX(1,2)D-mediated endocytosis. J Cell Biol. 2002; 157(2):315-26. PMC: 2199253. DOI: 10.1083/jcb.200110027. View

Henry K, DHondt K, Chang J, Newpher T, Huang K, Hudson R . Scd5p and clathrin function are important for cortical actin organization, endocytosis, and localization of sla2p in yeast. Mol Biol Cell. 2002; 13(8):2607-25. PMC: 117929. DOI: 10.1091/mbc.e02-01-0012. View

Desrivieres S, Cooke F, Morales-Johansson H, Parker P, Hall M . Calmodulin controls organization of the actin cytoskeleton via regulation of phosphatidylinositol (4,5)-bisphosphate synthesis in Saccharomyces cerevisiae. Biochem J. 2002; 366(Pt 3):945-51. PMC: 1222839. DOI: 10.1042/BJ20020429. View

Yin H, Janmey P . Phosphoinositide regulation of the actin cytoskeleton. Annu Rev Physiol. 2002; 65:761-89. DOI: 10.1146/annurev.physiol.65.092101.142517. View

10.

Lee W, Oberle J, Cooper J . The role of the lissencephaly protein Pac1 during nuclear migration in budding yeast. J Cell Biol. 2003; 160(3):355-64. PMC: 2172672. DOI: 10.1083/jcb.200209022. View

11.

Aguilar R, Watson H, Wendland B . The yeast Epsin Ent1 is recruited to membranes through multiple independent interactions. J Biol Chem. 2003; 278(12):10737-43. DOI: 10.1074/jbc.M211622200. View

12.

Gourlay C, Dewar H, Warren D, Costa R, Satish N, Ayscough K . An interaction between Sla1p and Sla2p plays a role in regulating actin dynamics and endocytosis in budding yeast. J Cell Sci. 2003; 116(Pt 12):2551-64. DOI: 10.1242/jcs.00454. View

13.

Padron D, Wang Y, Yamamoto M, Yin H, Roth M . Phosphatidylinositol phosphate 5-kinase Ibeta recruits AP-2 to the plasma membrane and regulates rates of constitutive endocytosis. J Cell Biol. 2003; 162(4):693-701. PMC: 2173809. DOI: 10.1083/jcb.200302051. View

14.

Sekiya-Kawasaki M, Groen A, Cope M, Kaksonen M, Watson H, Zhang C . Dynamic phosphoregulation of the cortical actin cytoskeleton and endocytic machinery revealed by real-time chemical genetic analysis. J Cell Biol. 2003; 162(5):765-72. PMC: 2172809. DOI: 10.1083/jcb.200305077. View

15.

Kaksonen M, Sun Y, Drubin D . A pathway for association of receptors, adaptors, and actin during endocytic internalization. Cell. 2003; 115(4):475-87. DOI: 10.1016/s0092-8674(03)00883-3. View

16.

Verstreken P, Koh T, Schulze K, Zhai R, Hiesinger P, Zhou Y . Synaptojanin is recruited by endophilin to promote synaptic vesicle uncoating. Neuron. 2003; 40(4):733-48. DOI: 10.1016/s0896-6273(03)00644-5. View

17.

Merrifield C . Seeing is believing: imaging actin dynamics at single sites of endocytosis. Trends Cell Biol. 2004; 14(7):352-8. DOI: 10.1016/j.tcb.2004.05.008. View

18.

Jonsdottir G, Li R . Dynamics of yeast Myosin I: evidence for a possible role in scission of endocytic vesicles. Curr Biol. 2004; 14(17):1604-9. DOI: 10.1016/j.cub.2004.08.055. View

19.

Paolo G, Moskowitz H, Gipson K, Wenk M, Voronov S, Obayashi M . Impaired PtdIns(4,5)P2 synthesis in nerve terminals produces defects in synaptic vesicle trafficking. Nature. 2004; 431(7007):415-22. DOI: 10.1038/nature02896. View

20.

Vida T, Emr S . A new vital stain for visualizing vacuolar membrane dynamics and endocytosis in yeast. J Cell Biol. 1995; 128(5):779-92. PMC: 2120394. DOI: 10.1083/jcb.128.5.779. View