Genetic Deletion of ABP-120 Alters the Three-dimensional Organization of Actin Filaments in Dictyostelium Pseudopods

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 1995 Mar 1

PMID 7876307

Citations 20

Authors

D Cox

J A Ridsdale

J Condeelis

J Hartwig

Affiliations

Soon will be listed here.

Abstract

This study extends the observations on the defects in pseudopod formation of ABP-120+ and ABP-120- cells by a detailed morphological and biochemical analysis of the actin based cytoskeleton. Both ABP-120+ and ABP-120- cells polymerize the same amount of F-actin in response to stimulation with cAMP. However, unlike ABP-120+ cells, ABP-120- cells do not incorporate actin into the Triton X-100-insoluble cytoskeleton at 30-50 s, the time when ABP-120 is incorporated into the cytoskeleton and when pseudopods are extended after cAMP stimulation in wild-type cells. By confocal and electron microscopy, pseudopods extended by ABP-120- cells are not as large or thick as those produced by ABP-120+ cells and in the electron microscope, an altered filament network is found in pseudopods of ABP-120- cells when compared to pseudopods of ABP-120+ cells. The actin filaments found in areas of pseudopods in ABP-120+ cells either before or after stimulation were long, straight, and arranged into space filling orthogonal networks. Protrusions of ABP-120- cells are less three-dimensional, denser, and filled with multiple foci of aggregated filaments consistent with collapse of the filament network due to the absence of ABP-120-mediated cross-linking activity. The different organization of actin filaments may account for the diminished size of protrusions observed in living and fixed ABP-120- cells compared to ABP-120+ cells and is consistent with the role of ABP-120 in regulating pseudopod extension through its cross-linking of actin filaments.

Citing Articles

Regulation of the Actin Cytoskeleton via Rho GTPase Signalling in and Mammalian Cells: A Parallel Slalom.

Filic V, Mijanovic L, Putar D, Talajic A, Cetkovic H, Weber I Cells. 2021; 10(7).

PMID: 34202767 PMC: 8305917. DOI: 10.3390/cells10071592.

Mechanosensitive Adhesion Explains Stepping Motility in Amoeboid Cells.

Copos C, Walcott S, Del Alamo J, Bastounis E, Mogilner A, Guy R Biophys J. 2017; 112(12):2672-2682.

PMID: 28636923 PMC: 5478966. DOI: 10.1016/j.bpj.2017.04.033.

Three-dimensional balance of cortical tension and axial contractility enables fast amoeboid migration.

Alvarez-Gonzalez B, Meili R, Bastounis E, Firtel R, Lasheras J, Del Alamo J Biophys J. 2015; 108(4):821-832.

PMID: 25692587 PMC: 4336364. DOI: 10.1016/j.bpj.2014.11.3478.

Both contractile axial and lateral traction force dynamics drive amoeboid cell motility.

Bastounis E, Meili R, Alvarez-Gonzalez B, Francois J, Del Alamo J, Firtel R J Cell Biol. 2014; 204(6):1045-61.

PMID: 24637328 PMC: 3998796. DOI: 10.1083/jcb.201307106.

Cease-fire at the leading edge: new perspectives on actin filament branching, debranching, and cross-linking.

Ydenberg C, Smith B, Breitsprecher D, Gelles J, Goode B Cytoskeleton (Hoboken). 2011; 68(11):596-602.

PMID: 22002930 PMC: 3218376. DOI: 10.1002/cm.20543.

References

Condeelis J . Life at the leading edge: the formation of cell protrusions. Annu Rev Cell Biol. 1993; 9:411-44. DOI: 10.1146/annurev.cb.09.110193.002211. View

Wessels D, Murray J, Jung G, Hammer 3rd J, Soll D . Myosin IB null mutants of Dictyostelium exhibit abnormalities in motility. Cell Motil Cytoskeleton. 1991; 20(4):301-15. DOI: 10.1002/cm.970200406. View

Omann G, Allen R, Bokoch G, Painter R, Traynor A, Sklar L . Signal transduction and cytoskeletal activation in the neutrophil. Physiol Rev. 1987; 67(1):285-322. DOI: 10.1152/physrev.1987.67.1.285. View

Condeelis J . Understanding the cortex of crawling cells: insights from Dictyostelium. Trends Cell Biol. 1993; 3(11):371-6. DOI: 10.1016/0962-8924(93)90085-f. View

Condeelis J, Bresnick A, Demma M, Dharmawardhane S, Eddy R, Hall A . Mechanisms of amoeboid chemotaxis: an evaluation of the cortical expansion model. Dev Genet. 1990; 11(5-6):333-40. DOI: 10.1002/dvg.1020110504. View

Hartwig J, Shevlin P . The architecture of actin filaments and the ultrastructural location of actin-binding protein in the periphery of lung macrophages. J Cell Biol. 1986; 103(3):1007-20. PMC: 2114287. DOI: 10.1083/jcb.103.3.1007. View

Hock R, Condeelis J . Isolation of a 240-kilodalton actin-binding protein from Dictyostelium discoideum. J Biol Chem. 1987; 262(1):394-400. View

Condeelis J, Vahey M, Carboni J, Demey J, Ogihara S . Properties of the 120,000- and 95,000-dalton actin-binding proteins from Dictyostelium discoideum and their possible functions in assembling the cytoplasmic matrix. J Cell Biol. 1984; 99(1 Pt 2):119s-126s. PMC: 2275594. DOI: 10.1083/jcb.99.1.119s. View

Ogihara S, Carboni J, Condeelis J . Electron microscopic localization of myosin II and ABP-120 in the cortical actin matrix of Dictyostelium amoebae using IgG-gold conjugates. Dev Genet. 1988; 9(4-5):505-20. DOI: 10.1002/dvg.1020090427. View

10.

Hartwig J, Kwiatkowski D . Actin-binding proteins. Curr Opin Cell Biol. 1991; 3(1):87-97. DOI: 10.1016/0955-0674(91)90170-4. View

11.

Egelhoff T, Spudich J . Molecular genetics of cell migration: Dictyostelium as a model system. Trends Genet. 1991; 7(5):161-6. DOI: 10.1016/0168-9525(91)90380-9. View

12.

Gorlin J, Yamin R, Egan S, Stewart M, Stossel T, Kwiatkowski D . Human endothelial actin-binding protein (ABP-280, nonmuscle filamin): a molecular leaf spring. J Cell Biol. 1990; 111(3):1089-105. PMC: 2116286. DOI: 10.1083/jcb.111.3.1089. View

13.

Zigmond S . Recent quantitative studies of actin filament turnover during cell locomotion. Cell Motil Cytoskeleton. 1993; 25(4):309-16. DOI: 10.1002/cm.970250402. View

14.

Cooper J . The role of actin polymerization in cell motility. Annu Rev Physiol. 1991; 53:585-605. DOI: 10.1146/annurev.ph.53.030191.003101. View

15.

Hall A, Warren V, Dharmawardhane S, Condeelis J . Identification of actin nucleation activity and polymerization inhibitor in ameboid cells: their regulation by chemotactic stimulation. J Cell Biol. 1989; 109(5):2207-13. PMC: 2115889. DOI: 10.1083/jcb.109.5.2207. View

16.

Cox D, Condeelis J, Wessels D, Soll D, Kern H, Knecht D . Targeted disruption of the ABP-120 gene leads to cells with altered motility. J Cell Biol. 1992; 116(4):943-55. PMC: 2289347. DOI: 10.1083/jcb.116.4.943. View

17.

Titus M, Wessels D, Spudich J, Soll D . The unconventional myosin encoded by the myoA gene plays a role in Dictyostelium motility. Mol Biol Cell. 1993; 4(2):233-46. PMC: 300918. DOI: 10.1091/mbc.4.2.233. View

18.

Lewis A, Bridgman P . Nerve growth cone lamellipodia contain two populations of actin filaments that differ in organization and polarity. J Cell Biol. 1992; 119(5):1219-43. PMC: 2289720. DOI: 10.1083/jcb.119.5.1219. View

19.

Hartwig J, Yin H . The organization and regulation of the macrophage actin skeleton. Cell Motil Cytoskeleton. 1988; 10(1-2):117-25. DOI: 10.1002/cm.970100116. View

20.

Fukui Y . Toward a new concept of cell motility: cytoskeletal dynamics in amoeboid movement and cell division. Int Rev Cytol. 1993; 144:85-127. DOI: 10.1016/s0074-7696(08)61514-4. View