Actin Pedestal Formation by Enteropathogenic Escherichia Coli and Intracellular Motility of Shigella Flexneri Are Abolished in N-WASP-defective Cells

Overview

Journal EMBO Rep

Specialty Molecular Biology

Date 2001 Sep 18

PMID 11559594

Citations 100

Authors

S Lommel

S Benesch

K Rottner

T Franz

J Wehland

R Kuhn

Affiliations

Soon will be listed here.

Abstract

In mammalian cells, actin dynamics is tightly controlled through small GTPases of the Rho family, WASP/Scar proteins and the Arp2/3 complex. We employed Cre/loxP-mediated gene targeting to disrupt the ubiquitously expressed N-WASP in the mouse germline, which led to embryonic lethality. To elucidate the role of N-WASP at the cellular level, we immortalized embryonic fibroblasts and selected various N-WASP-defective cell lines. These fibroblasts showed no apparent morphological alterations and were highly responsive to the induction of filopodia, but failed to support the motility of Shigella flexneri. In addition, enteropathogenic Escherichia coli were incapable of inducing the formation of actin pedestals in N-WASP-defective cells. Our results prove the essential role of this protein for actin cytoskeletal changes induced by these bacterial pathogens in vivo and in addition show for the first time that N-WASP is dispensable for filopodia formation.

Citing Articles

Nuclear N-WASP Induces Actin Polymerization in the Nucleus with Cortactin as an Essential Factor.

Jiang X, Mohapatra P, Rossing M, Zheng W, Zbodakova O, Thatte J Cells. 2025; 14(1.

PMID: 39791760 PMC: 11720165. DOI: 10.3390/cells14010059.

Cytoskeletal mechanisms regulating attaching/effacing bacteria interactions with host cells: It takes a village to build the pedestal.

Naydenov N, Marino-Melendez A, Campellone K, Ivanov A Bioessays. 2024; 46(11):e2400160.

PMID: 39301984 PMC: 11502255. DOI: 10.1002/bies.202400160.

Branching out in different directions: Emerging cellular functions for the Arp2/3 complex and WASP-family actin nucleation factors.

Campellone K, Lebek N, King V Eur J Cell Biol. 2023; 102(2):151301.

PMID: 36907023 PMC: 10330494. DOI: 10.1016/j.ejcb.2023.151301.

Lamellipodia-like actin networks in cells lacking WAVE regulatory complex.

Kage F, Doring H, Mietkowska M, Schaks M, Gruner F, Stahnke S J Cell Sci. 2022; 135(15).

PMID: 35971979 PMC: 9511706. DOI: 10.1242/jcs.260364.

WASP family proteins: Molecular mechanisms and implications in human disease.

Kramer D, K Piper H, Chen B Eur J Cell Biol. 2022; 101(3):151244.

PMID: 35667337 PMC: 9357188. DOI: 10.1016/j.ejcb.2022.151244.

References

Frischknecht F, Moreau V, Rottger S, Gonfloni S, Reckmann I, Superti-Furga G . Actin-based motility of vaccinia virus mimics receptor tyrosine kinase signalling. Nature. 1999; 401(6756):926-9. DOI: 10.1038/44860. View

Kalman D, Weiner O, Goosney D, Sedat J, Finlay B, Abo A . Enteropathogenic E. coli acts through WASP and Arp2/3 complex to form actin pedestals. Nat Cell Biol. 1999; 1(6):389-91. PMC: 2828053. DOI: 10.1038/14087. View

Minichiello L, Korte M, Wolfer D, Kuhn R, Unsicker K, Cestari V . Essential role for TrkB receptors in hippocampus-mediated learning. Neuron. 1999; 24(2):401-14. DOI: 10.1016/s0896-6273(00)80853-3. View

Suzuki T, Mimuro H, Miki H, Takenawa T, Sasaki T, Nakanishi H . Rho family GTPase Cdc42 is essential for the actin-based motility of Shigella in mammalian cells. J Exp Med. 2000; 191(11):1905-20. PMC: 2213524. DOI: 10.1084/jem.191.11.1905. View

Egile C, Loisel T, Laurent V, Li R, Pantaloni D, Sansonetti P . Activation of the CDC42 effector N-WASP by the Shigella flexneri IcsA protein promotes actin nucleation by Arp2/3 complex and bacterial actin-based motility. J Cell Biol. 1999; 146(6):1319-32. PMC: 2156126. DOI: 10.1083/jcb.146.6.1319. View

Loisel T, Boujemaa R, Pantaloni D, Carlier M . Reconstitution of actin-based motility of Listeria and Shigella using pure proteins. Nature. 1999; 401(6753):613-6. DOI: 10.1038/44183. View

Moreau V, Frischknecht F, Reckmann I, Vincentelli R, Rabut G, Stewart D . A complex of N-WASP and WIP integrates signalling cascades that lead to actin polymerization. Nat Cell Biol. 2000; 2(7):441-8. DOI: 10.1038/35017080. View

Carlier M, Nioche P, Boujemaa R, Le Clainche C, Egile C, Garbay C . GRB2 links signaling to actin assembly by enhancing interaction of neural Wiskott-Aldrich syndrome protein (N-WASp) with actin-related protein (ARP2/3) complex. J Biol Chem. 2000; 275(29):21946-52. DOI: 10.1074/jbc.M000687200. View

Mimuro H, Suzuki T, Suetsugu S, Miki H, Takenawa T, Sasakawa C . Profilin is required for sustaining efficient intra- and intercellular spreading of Shigella flexneri. J Biol Chem. 2000; 275(37):28893-901. DOI: 10.1074/jbc.M003882200. View

10.

Goosney D, Gruenheid S, Finlay B . Gut feelings: enteropathogenic E. coli (EPEC) interactions with the host. Annu Rev Cell Dev Biol. 2000; 16:173-89. DOI: 10.1146/annurev.cellbio.16.1.173. View

11.

Prehoda K, Scott J, Mullins R, Lim W . Integration of multiple signals through cooperative regulation of the N-WASP-Arp2/3 complex. Science. 2000; 290(5492):801-6. DOI: 10.1126/science.290.5492.801. View

12.

Yamaguchi H, Miki H, Suetsugu S, Ma L, Kirschner M, Takenawa T . Two tandem verprolin homology domains are necessary for a strong activation of Arp2/3 complex-induced actin polymerization and induction of microspike formation by N-WASP. Proc Natl Acad Sci U S A. 2000; 97(23):12631-6. PMC: 18815. DOI: 10.1073/pnas.190351397. View

13.

Frischknecht F, Way M . Surfing pathogens and the lessons learned for actin polymerization. Trends Cell Biol. 2001; 11(1):30-38. DOI: 10.1016/s0962-8924(00)01871-7. View

14.

Erickson J, Cerione R . Multiple roles for Cdc42 in cell regulation. Curr Opin Cell Biol. 2001; 13(2):153-7. DOI: 10.1016/s0955-0674(00)00192-7. View

15.

Goosney D, DeVinney R, Finlay B . Recruitment of cytoskeletal and signaling proteins to enteropathogenic and enterohemorrhagic Escherichia coli pedestals. Infect Immun. 2001; 69(5):3315-22. PMC: 98290. DOI: 10.1128/IAI.69.5.3315-3322.2001. View

16.

Takenawa T, Miki H . WASP and WAVE family proteins: key molecules for rapid rearrangement of cortical actin filaments and cell movement. J Cell Sci. 2001; 114(Pt 10):1801-9. DOI: 10.1242/jcs.114.10.1801. View

17.

Jat P, Sharp P . Cell lines established by a temperature-sensitive simian virus 40 large-T-antigen gene are growth restricted at the nonpermissive temperature. Mol Cell Biol. 1989; 9(4):1672-81. PMC: 362586. DOI: 10.1128/mcb.9.4.1672-1681.1989. View

18.

Domann E, Wehland J, Rohde M, Pistor S, Hartl M, Goebel W . A novel bacterial virulence gene in Listeria monocytogenes required for host cell microfilament interaction with homology to the proline-rich region of vinculin. EMBO J. 1992; 11(5):1981-90. PMC: 556658. DOI: 10.1002/j.1460-2075.1992.tb05252.x. View

19.

Kozma R, Ahmed S, Best A, Lim L . The Ras-related protein Cdc42Hs and bradykinin promote formation of peripheral actin microspikes and filopodia in Swiss 3T3 fibroblasts. Mol Cell Biol. 1995; 15(4):1942-52. PMC: 230420. DOI: 10.1128/MCB.15.4.1942. View

20.

Nobes C, Hall A . Rho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia. Cell. 1995; 81(1):53-62. DOI: 10.1016/0092-8674(95)90370-4. View