Active Arf6 Recruits ARNO/cytohesin GEFs to the PM by Binding Their PH Domains

Overview

Journal Mol Biol Cell

Specialties Cell Biology
Molecular Biology

Date 2007 Apr 6

PMID 17409355

Citations 135

Authors

Lee Ann Cohen

Akira Honda

Peter Varnai

Fraser D Brown

Tamas Balla

Julie G Donaldson

Affiliations

Soon will be listed here.

Abstract

ARNO is a soluble guanine nucleotide exchange factor (GEF) for the Arf family of GTPases. Although in biochemical assays ARNO prefers Arf1 over Arf6 as a substrate, its localization in cells at the plasma membrane (PM) suggests an interaction with Arf6. In this study, we found that ARNO activated Arf1 in HeLa and COS-7 cells resulting in the recruitment of Arf1 on to dynamic PM ruffles. By contrast, Arf6 was activated less by ARNO than EFA6, a canonical Arf6 GEF. Remarkably, Arf6 in its GTP-bound form recruited ARNO to the PM and the two proteins could be immunoprecipitated. ARNO binding to Arf6 was not mediated through the catalytic Sec7 domain, but via the pleckstrin homology (PH) domain. Active Arf6 also bound the PH domain of Grp1, another ARNO family member. This interaction was direct and required both inositol phospholipids and GTP. We propose a model of sequential Arf activation at the PM whereby Arf6-GTP recruits ARNO family GEFs for further activation of other Arf isoforms.

Citing Articles

Single-molecule lipid biosensors mitigate inhibition of endogenous effector proteins.

Holmes V, Ricci M, Weckerly C, Worcester M, Hammond G J Cell Biol. 2025; 224(3).

PMID: 39932556 PMC: 11812570. DOI: 10.1083/jcb.202412026.

Single molecule Lipid Biosensors Mitigate Inhibition of Endogenous Effector Proteins.

Holmes V, Ricci M, Weckerly C, Worcester M, Hammond G bioRxiv. 2024; .

PMID: 39345595 PMC: 11429874. DOI: 10.1101/2024.09.11.612480.

Arf6 as a therapeutic target: Structure, mechanism, and inhibitors.

Sun D, Guo Y, Tang P, Li H, Chen L Acta Pharm Sin B. 2023; 13(10):4089-4104.

PMID: 37799386 PMC: 10547916. DOI: 10.1016/j.apsb.2023.06.008.

The Arf-GAP Age2 localizes to the late-Golgi via a conserved amphipathic helix.

Manzer K, Fromme J Mol Biol Cell. 2023; 34(12):ar119.

PMID: 37672345 PMC: 10846627. DOI: 10.1091/mbc.E23-07-0283.

The Arf-GAP Age2 localizes to the late-Golgi via a conserved amphipathic helix.

Manzer K, Fromme J bioRxiv. 2023; .

PMID: 37546741 PMC: 10402032. DOI: 10.1101/2023.07.23.550229.

References

Santy L, Casanova J . Activation of ARF6 by ARNO stimulates epithelial cell migration through downstream activation of both Rac1 and phospholipase D. J Cell Biol. 2001; 154(3):599-610. PMC: 2196419. DOI: 10.1083/jcb.200104019. View

Varnai P, Bondeva T, Tamas P, Toth B, Buday L, Hunyady L . Selective cellular effects of overexpressed pleckstrin-homology domains that recognize PtdIns(3,4,5)P3 suggest their interaction with protein binding partners. J Cell Sci. 2005; 118(Pt 20):4879-88. DOI: 10.1242/jcs.02606. View

Presley J, Ward T, Pfeifer A, Siggia E, Phair R, Lippincott-Schwartz J . Dissection of COPI and Arf1 dynamics in vivo and role in Golgi membrane transport. Nature. 2002; 417(6885):187-93. DOI: 10.1038/417187a. View

Campbell R, Tour O, Palmer A, Steinbach P, Baird G, Zacharias D . A monomeric red fluorescent protein. Proc Natl Acad Sci U S A. 2002; 99(12):7877-82. PMC: 122988. DOI: 10.1073/pnas.082243699. View

Hernandez-Deviez D, Casanova J, Wilson J . Regulation of dendritic development by the ARF exchange factor ARNO. Nat Neurosci. 2002; 5(7):623-4. DOI: 10.1038/nn865. View

Varnai P, Lin X, Lee S, Tuymetova G, Bondeva T, Spat A . Inositol lipid binding and membrane localization of isolated pleckstrin homology (PH) domains. Studies on the PH domains of phospholipase C delta 1 and p130. J Biol Chem. 2002; 277(30):27412-22. DOI: 10.1074/jbc.M109672200. View

Mariana Margarit S, Sondermann H, Hall B, Nagar B, Hoelz A, Pirruccello M . Structural evidence for feedback activation by Ras.GTP of the Ras-specific nucleotide exchange factor SOS. Cell. 2003; 112(5):685-95. DOI: 10.1016/s0092-8674(03)00149-1. View

Nie Z, Hirsch D, Randazzo P . Arf and its many interactors. Curr Opin Cell Biol. 2003; 15(4):396-404. DOI: 10.1016/s0955-0674(03)00071-1. View

Cheng L, Mahon G, Kostenko E, Whitehead I . Pleckstrin homology domain-mediated activation of the rho-specific guanine nucleotide exchange factor Dbs by Rac1. J Biol Chem. 2004; 279(13):12786-93. DOI: 10.1074/jbc.M313099200. View

10.

Santy L, Frank S, Hatfield J, Casanova J . Regulation of ARNO nucleotide exchange by a PH domain electrostatic switch. Curr Biol. 1999; 9(20):1173-6. DOI: 10.1016/S0960-9822(00)80019-6. View

11.

Honda A, Nogami M, Yokozeki T, Yamazaki M, Nakamura H, Watanabe H . Phosphatidylinositol 4-phosphate 5-kinase alpha is a downstream effector of the small G protein ARF6 in membrane ruffle formation. Cell. 1999; 99(5):521-32. DOI: 10.1016/s0092-8674(00)81540-8. View

12.

Ogasawara M, Kim S, Adamik R, Togawa A, Ferrans V, Takeda K . Similarities in function and gene structure of cytohesin-4 and cytohesin-1, guanine nucleotide-exchange proteins for ADP-ribosylation factors. J Biol Chem. 2000; 275(5):3221-30. DOI: 10.1074/jbc.275.5.3221. View

13.

Jackson C, Casanova J . Turning on ARF: the Sec7 family of guanine-nucleotide-exchange factors. Trends Cell Biol. 2000; 10(2):60-7. DOI: 10.1016/s0962-8924(99)01699-2. View

14.

Rozelle A, Machesky L, Yamamoto M, Driessens M, Insall R, Roth M . Phosphatidylinositol 4,5-bisphosphate induces actin-based movement of raft-enriched vesicles through WASP-Arp2/3. Curr Biol. 2000; 10(6):311-20. DOI: 10.1016/s0960-9822(00)00384-5. View

15.

Menetrey J, Macia E, Pasqualato S, Franco M, CHERFILS J . Structure of Arf6-GDP suggests a basis for guanine nucleotide exchange factors specificity. Nat Struct Biol. 2000; 7(6):466-9. DOI: 10.1038/75863. View

16.

Godi A, Santone I, Pertile P, Marra P, Di Tullio G, Luini A . ADP-ribosylation factor regulates spectrin skeleton assembly on the Golgi complex by stimulating phosphatidylinositol 4,5-bisphosphate synthesis. Biochem Soc Trans. 2000; 27(4):638-42. DOI: 10.1042/bst0270638. View

17.

Klarlund J, Tsiaras W, Holik J, Chawla A, Czech M . Distinct polyphosphoinositide binding selectivities for pleckstrin homology domains of GRP1-like proteins based on diglycine versus triglycine motifs. J Biol Chem. 2000; 275(42):32816-21. DOI: 10.1074/jbc.M002435200. View

18.

Someya A, Sata M, Takeda K, Ferrans V, Moss J, Vaughan M . ARF-GEP(100), a guanine nucleotide-exchange protein for ADP-ribosylation factor 6. Proc Natl Acad Sci U S A. 2001; 98(5):2413-8. PMC: 30152. DOI: 10.1073/pnas.051634798. View

19.

Pasqualato S, Menetrey J, Franco M, CHERFILS J . The structural GDP/GTP cycle of human Arf6. EMBO Rep. 2001; 2(3):234-8. PMC: 1083839. DOI: 10.1093/embo-reports/kve043. View

20.

Maranda B, Brown D, Bourgoin S, Casanova J, Vinay P, Ausiello D . Intra-endosomal pH-sensitive recruitment of the Arf-nucleotide exchange factor ARNO and Arf6 from cytoplasm to proximal tubule endosomes. J Biol Chem. 2001; 276(21):18540-50. DOI: 10.1074/jbc.M011577200. View