Cross-linking of actin filaments by myosin II is a major contributor to cortical integrity and cell motility in restrictive environments
Abstract
Cells are frequently required to move in a local environment that physically restricts locomotion, such as during extravasation or metastatic invasion. In order to model these events, we have developed an assay in which vegetative Dictyostelium amoebae undergo chemotaxis under a layer of agarose toward a source of folic acid [Laevsky, G. and Knecht, D. A. (2001). Biotechniques 31, 1140-1149]. As the concentration of agarose is increased from 0.5% to 3% the cells are increasingly inhibited in their ability to move under the agarose. The contribution of myosin II and actin cross-linking proteins to the movement of cells in this restrictive environment has now been examined. Cells lacking myosin II heavy chain (mhcA-) are unable to migrate under agarose overlays of greater than 0.5%, and even at this concentration they move only a short distance from the trough. While attempting to move, the cells become stretched and fragmented due to their inability to retract their uropods. At higher agarose concentrations, the mhcA- cells protrude pseudopods under the agarose, but are unable to pull the cell body underneath. Consistent with a role for myosin II in general cortical stability, GFP-myosin dynamically localizes to the lateral and posterior cortex of cells moving under agarose. Cells lacking the essential light chain of myosin II (mlcE-), have no measurable myosin II motor activity, yet were able to move normally under all agarose concentrations. Mutants lacking either ABP-120 or α-actinin were also able to move under agarose at rates similar to wild-type cells. We hypothesize that myosin stabilizes the actin cortex through its cross-linking activity rather than its motor function and this activity is necessary and sufficient for the maintenance of cortical integrity of cells undergoing movement in a restrictive environment. The actin cross-linkers α-actinin and ABP-120 do not appear to play as major a role as myosin II in providing this cortical integrity.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Stress filaments consist of actin, myosin-II dimers, and associated crosslinkers.
“Stress filaments consist of actin, myosin-II dimers, and associated crosslinkers (Fig. 3c) [32–34].”
From Macrophages bend long fibres with flexural rigidity lower than 3 mN·nm2 to avoid frustrated phagocytosis (2026) · cites this paper as [32]
Cites
G Protein Signaling Events Are Activated at the Leading Edge of Chemotactic Cells
Parent, C. A., Blacklock, B. J., Froehlich, W. M., Murphy, D. B., & Devreotes, P. N. (1998) · Cell · doi:10.1016/s0092-8674(00)81784-5 · Cited by 617 · see citing works
The acrasin activity of adenosine-3',5'-cyclic phosphate.
Konijn, T. M., Jgc, V. D. M., Bonner, J. T., et al. (1967) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.58.3.1152 · Cited by 503 · see citing works
Active fluidization of polymer networks through molecular motors
Humphrey, D., Duggan, C., Saha, D., Smith, D. W., & Käs, J. A. (2002) · Nature · doi:10.1038/416413a · Cited by 328 · see citing works
Cell motility and chemotaxis in Dictyostelium amebae lacking myosin heavy chain
Wessels, D. J., Soll, D. R., Knecht, D. A., Loomis, W. F., Lozanne, A. D., & Spudich, J. A. (1988) · Developmental Biology · doi:10.1016/0012-1606(88)90279-5 · Cited by 326 · see citing works
Self-citationYumura, S., Mori, H., & Fukui, Y. (1984) · The Journal of Cell Biology · doi:10.1083/jcb.99.3.894 · Cited by 310 · see citing works
Acrasin, acrasinase, and the sensitivity to acrasin in Dictyostelium discoideum
Bonner, J. T., Barkley, D. S., Hall, E. M., et al. (1969) · Developmental Biology · doi:10.1016/0012-1606(69)90005-0 · Cited by 301 · see citing works
Egelhoff, T. T., Lee, R. J., & Spudich, J. A. (1993) · Cell · doi:10.1016/0092-8674(93)80077-r · Cited by 282 · see citing works
Capping of surface receptors and concomitant cortical tension are generated by conventional myosin
Pasternak, C., Spudich, J. A., & Elson, E. L. (1989) · Nature · doi:10.1038/341549a0 · Cited by 272 · see citing works
A mechanical function of myosin II in cell motility
Jay, P. Y., Pham, P. A., Wong, S. A., & Elson, E. L. (1995) · Journal of Cell Science · doi:10.1242/jcs.108.1.387 · Cited by 259 · see citing works
Cross-linker dynamics determine the mechanical properties of actin gels
Wachsstock, D. H., Schwarz, W. H., & Pollard, T. D. (1994) · Biophysical Journal · doi:10.1016/s0006-3495(94)80856-2 · Cited by 249 · see citing works
Biochemical and structural studies of actomyosin-like proteins from non-muscle cells
Clarke, M., & Spudich, J. A. (1974) · Journal of Molecular Biology · doi:10.1016/0022-2836(74)90013-8 · Cited by 231 · see citing works
A Cellophane-Strip Technique for Culturing Tissue in Multipurpose Culture Chambers
Rose, G. G., POMERAT, C. M., Shindler, T. O., et al. (1958) · The Journal of Cell Biology · doi:10.1083/jcb.4.6.761 · Cited by 220 · see citing works
Myosin dynamics in live Dictyostelium cells.
Moores, S. L., Sabry, J. H., & Spudich, J. A. (1996) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.93.1.443 · Cited by 205 · see citing works
Faix, J., Steinmetz, M. O., Boves, H., Kammerer, R. A., Lottspeich, F., Mintert, U., Murphy, J., Stock, A., Aebi, U., & Gerisch, G. (1996) · Cell · doi:10.1016/s0092-8674(00)80136-1 · Cited by 176 · see citing works
Neujahr, R., Heizer, C., & Gerisch, G. (1997) · Journal of Cell Science · doi:10.1242/jcs.110.2.123 · Cited by 169 · see citing works
Pang, K. M., Lee, E., & Knecht, D. A. (1998) · Current Biology · doi:10.1016/s0960-9822(98)70159-9 · Cited by 135 · see citing works
Self-citationCultivation of Dictyostelium discoideum in axenic medium
Sussman, R., & Sussman, M. (1967) · Biochemical and Biophysical Research Communications · doi:10.1016/0006-291x(67)90539-6 · Cited by 133 · see citing works
Noegel, A. A., Rapp, S., Lottspeich, F., et al. (1989) · The Journal of Cell Biology · doi:10.1083/jcb.109.2.607 · Cited by 130 · see citing works
A calcium- and pH-regulated protein from Dictyostelium discoideum that cross-links actin filaments.
Condeelis, J. S., & Vahey, M. (1982) · The Journal of Cell Biology · doi:10.1083/jcb.94.2.466 · Cited by 129 · see citing works
Targeted disruption of the ABP-120 gene leads to cells with altered motility.
Cox, D., Condeelis, J. S., Wessels, D. J., et al. (1992) · The Journal of Cell Biology · doi:10.1083/jcb.116.4.943 · Cited by 124 · see citing works
A Micromechanic Study of Cell Polarity and Plasma Membrane Cell Body Coupling in Dictyostelium
Merkel, R., Simson, R., Simson, D. A., et al. (2000) · Biophysical Journal · doi:10.1016/s0006-3495(00)76329-6 · Cited by 117 · see citing works
Griffith, L. M., Downs, S. M., & Spudich, J. A. (1987) · The Journal of Cell Biology · doi:10.1083/jcb.104.5.1309 · Cited by 117 · see citing works
Condeelis, J. S., Vahey, M., Carboni, J. M., et al. (1984) · The Journal of Cell Biology · doi:10.1083/jcb.99.1.119s · Cited by 104 · see citing works
Dictyostelium cells' cytoplasm as an active viscoplastic body
Feneberg, W., Westphal, M., & Sackmann, E. (2001) · European Biophysics Journal · doi:10.1007/s002490100135 · Cited by 94 · see citing works
Movement of myosin fragments in vitro: Domains involved in force production
Hynes, T. R., Block, S. M., White, B. T., & Spudich, J. A. (1987) · Cell · doi:10.1016/0092-8674(87)90704-5 · Cited by 89 · see citing works
Adenosine-3′,5′-Phosphate: Identification as Acrasin in a Species of Cellular Slime Mold
Barkley, D. S. (1969) · Science · doi:10.1126/science.165.3898.1133 · Cited by 85 · see citing works
Prassler, J., Stocker, S., Marriott, G., et al. (1997) · Molecular Biology of the Cell · doi:10.1091/mbc.8.1.83 · Cited by 82 · see citing works
Fechheimer, M., & Taylor, D. L. (1984) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)43076-6 · Cited by 74 · see citing works
Laevsky, G. S., & Knecht, D. A. (2001) · BioTechniques · doi:10.2144/01315rr03 · Cited by 72 · see citing works
Self-citationMutants lacking myosin II cannot resist forces generated during multicellular morphogenesis
Shelden, E. A., & Knecht, D. A. (1995) · Journal of Cell Science · doi:10.1242/jcs.108.3.1105 · Cited by 72 · see citing works
Self-citation[50] Myosin movement in Vitro: A quantitative assay using oriented actin cables from Nitella
Sheetz, M. P., Block, S. M., & Spudich, J. A. (1986) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/0076-6879(86)34118-1 · Cited by 70 · see citing works
Developmental consequences of the lack of myosin heavy chain in Dictyostelium discoideum
Knecht, D. A., & Loomis, W. F. (1988) · Developmental Biology · doi:10.1016/0012-1606(88)90280-1 · Cited by 69 · see citing works
Self-citationThe dictyostelium essential light chain is required for myosin function
Pollenz, R. S., Chen, T.-L. L., Triviños-Lagos, L., et al. (1992) · Cell · doi:10.1016/0092-8674(92)90614-i · Cited by 68 · see citing works
In Vivo Observations of Myosin II Dynamics Support a Role in Rear Retraction
Clow, P. A., & McNally, J. G. (1999) · Molecular Biology of the Cell · doi:10.1091/mbc.10.5.1309 · Cited by 68 · see citing works
Rivero, F., Furukawa, R., Fechheimer, M., & Noegel, A. A. (1999) · Journal of Cell Science · doi:10.1242/jcs.112.16.2737 · Cited by 67 · see citing works
Chen, T.-L. L., Kowalczyk, P. A., Ho, G., et al. (1995) · Journal of Cell Science · doi:10.1242/jcs.108.10.3207 · Cited by 64 · see citing works
A calcium‐ and pH‐regulated actin binding protein from D. discoideum
Fechheimer, M., Brier, J., Rockwell, M., et al. (1982) · Cell Motility · doi:10.1002/cm.970020308 · Cited by 63 · see citing works
Cox, D., Wessels, D. J., Soll, D. R., Hartwig, J. H., & Condeelis, J. S. (1996) · Molecular Biology of the Cell · doi:10.1091/mbc.7.5.803 · Cited by 62 · see citing works
Peters, D. J. M., Knecht, D. A., Loomis, W. F., et al. (1988) · Developmental Biology · doi:10.1016/0012-1606(88)90278-3 · Cited by 51 · see citing works
Self-citationDuring Multicellular Migration, Myosin II Serves a Structural Role Independent of its Motor Function
Xu, X., Lee, E., Chen, T.-L., et al. (2001) · Developmental Biology · doi:10.1006/dbio.2000.0132 · Cited by 49 · see citing works
A new protein that gels F actin in the cell cortex of Dictyostelium discoideum
Coudeelis, J., Salisbury, J., & Fujiwara, K. (1981) · Nature · doi:10.1038/292161a0 · Cited by 44 · see citing works
Egelhoff, T. T., Naismith, T. V., & Brozovich, F. V. (1996) · Journal of Muscle Research and Cell Motility · doi:10.1007/bf00124248 · Cited by 41 · see citing works
Knecht, D. A., & Shelden, E. A. (1995) · Developmental Biology · doi:10.1006/dbio.1995.1227 · Cited by 38 · see citing works
Self-citationLiu, X., Osherov, N., Yamashita, R. A., et al. (2001) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.161285698 · Cited by 38 · see citing works
Burden‐Gulley, S. M., & Lemmon, V. (1996) · Cell Motility and the Cytoskeleton · doi:10.1002/(sici)1097-0169(1996)35:1 · Cited by 35 · see citing works
Severe developmental defects in Dictyostelium null mutants for actin-binding proteins
Ponte, E., Rivero, F., Fechheimer, M., Noegel, A. A., & Bozzaro, S. (2000) · Mechanisms of Development · doi:10.1016/s0925-4773(99)00292-0 · Cited by 34 · see citing works
Regulation and Tuning of Smooth Muscle Myosin
Sweeney, H. L. (1998) · American Journal of Respiratory and Critical Care Medicine · doi:10.1164/ajrccm.158.supplement_2.13tac400 · Cited by 33 · see citing works
Xu, X., Kuspa, A., Fuller, D., et al. (1996) · Developmental Biology · doi:10.1006/dbio.1996.0109 · Cited by 27 · see citing works
A simple method of generating axenic derivatives of Dictyostelium strains
Morrison, A., & Harwood, A. J. (1992) · Experimental Cell Research · doi:10.1016/0014-4827(92)90449-i · Cited by 11 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Broßell, D., Meyer-Plath, A., Gräb, O., Heunisch, E., Kämpf, K., Haase, A., & Wiemann, M. (2026) · Particle and Fibre Toxicology · doi:10.1186/s12989-026-00666-9 · Cited by 2 · see citing works
- Authors
- Laevsky, G., & Knecht, D. A.
- Affiliations
- University of Connecticut (US)
- Corresponding author
- Laevsky, Gary
- Knecht, David A.
- Venue
- Journal of Cell Science
- Publisher
- The Company of Biologists
- Published
- 2003-09-15
- Volume
- 116
- Issue
- 18
- Pages
- 3761-3770
- ISSN
- 0021-9533 (ISSN-L)
- 1477-9137
- OpenAlex
- W2125509960
- Cited by
- 120
- References
- 49
- Citations by year
- 2026: 2 · 2025: 3 · 2024: 3 · 2023: 3 · 2022: 2 · 2021: 15 · 2020: 4 · 2019: 6 · 2018: 2 · 2017: 3 · 2016: 2 · 2015: 3 · 2014: 6 · 2013: 2 · 2012: 7
- FWCI
- 2.22
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Cell Science (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of General Medical Sciences (GM 40599, R01GM040599)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Keywords
- myosin II, actin cross-linking, actin cortex, cell motility, Dictyostelium, chemotaxis, myosin heavy chain, myosin light chain, alpha-actinin, restricted environments, metastatic invasion
- MeSH terms
- Actinin, Animals, Carrier Proteins, Cell Movement, Chemotaxis, Cross-Linking Reagents, Dictyostelium, Luminescent Proteins, Microfilament Proteins, Actin Cytoskeleton, Mutation, Recombinant Fusion Proteins, Sepharose, Myosin Light Chains, Myosin Type II, Green Fluorescent Proteins
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
