Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1994 Aug 2;126(4):1005–1015. doi: 10.1083/jcb.126.4.1005

Actin filament organization in activated mast cells is regulated by heterotrimeric and small GTP-binding proteins

PMCID: PMC2120121  PMID: 8051203

Abstract

Rat peritoneal mast cells, both intact and permeabilized, have been used widely as model secretory cells. GTP-binding proteins and calcium play a major role in controlling their secretory response. Here we have examined changes in the organization of actin filaments in intact mast cells after activation by compound 48/80, and in permeabilized cells after direct activation of GTP-binding proteins by GTP-gamma-S. In both cases, a centripetal redistribution of cellular F-actin was observed: the content of F-actin was reduced in the cortical region and increased in the cell interior. The overall F-actin content was increased. Using permeabilized cells, we show that AIF4-, an activator of heterotrimeric G proteins, induces the disassembly of F-actin at the cortex, while the appearance of actin filaments in the interior of the cell is dependent on two small GTPases, rho and rac. Rho was found to be responsible for de novo actin polymerization, presumably from a membrane-bound monomeric pool, while rac was required for an entrapment of the released cortical filaments. Thus, a heterotrimeric G-protein and the small GTPases, rho and rac, participate in affecting the changes in the actin cytoskeleton observed after activation of mast cells.

Full Text

The Full Text of this article is available as a PDF (1.6 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abo A., Pick E., Hall A., Totty N., Teahan C. G., Segal A. W. Activation of the NADPH oxidase involves the small GTP-binding protein p21rac1. Nature. 1991 Oct 17;353(6345):668–670. doi: 10.1038/353668a0. [DOI] [PubMed] [Google Scholar]
  2. Aktories K., Braun U., Rösener S., Just I., Hall A. The rho gene product expressed in E. coli is a substrate of botulinum ADP-ribosyltransferase C3. Biochem Biophys Res Commun. 1989 Jan 16;158(1):209–213. doi: 10.1016/s0006-291x(89)80199-8. [DOI] [PubMed] [Google Scholar]
  3. Aridor M., Rajmilevich G., Beaven M. A., Sagi-Eisenberg R. Activation of exocytosis by the heterotrimeric G protein Gi3. Science. 1993 Dec 3;262(5139):1569–1572. doi: 10.1126/science.7504324. [DOI] [PubMed] [Google Scholar]
  4. Aridor M., Traub L. M., Sagi-Eisenberg R. Exocytosis in mast cells by basic secretagogues: evidence for direct activation of GTP-binding proteins. J Cell Biol. 1990 Sep;111(3):909–917. doi: 10.1083/jcb.111.3.909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bar-Sagi D., Gomperts B. D. Stimulation of exocytotic degranulation by microinjection of the ras oncogene protein into rat mast cells. Oncogene. 1988 Oct;3(4):463–469. [PubMed] [Google Scholar]
  6. Bengtsson T., Särndahl E., Stendahl O., Andersson T. Involvement of GTP-binding proteins in actin polymerization in human neutrophils. Proc Natl Acad Sci U S A. 1990 Apr;87(8):2921–2925. doi: 10.1073/pnas.87.8.2921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bowman E. P., Uhlinger D. J., Lambeth J. D. Neutrophil phospholipase D is activated by a membrane-associated Rho family small molecular weight GTP-binding protein. J Biol Chem. 1993 Oct 15;268(29):21509–21512. [PubMed] [Google Scholar]
  8. Burgoyne R. D., Cheek T. R. Reorganisation of peripheral actin filaments as a prelude to exocytosis. Biosci Rep. 1987 Apr;7(4):281–288. doi: 10.1007/BF01121449. [DOI] [PubMed] [Google Scholar]
  9. Cao L. G., Fishkind D. J., Wang Y. L. Localization and dynamics of nonfilamentous actin in cultured cells. J Cell Biol. 1993 Oct;123(1):173–181. doi: 10.1083/jcb.123.1.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Carlson K. E., Woolkalis M. J., Newhouse M. G., Manning D. R. Fractionation of the beta subunit common to guanine nucleotide-binding regulatory proteins with the cytoskeleton. Mol Pharmacol. 1986 Nov;30(5):463–468. [PubMed] [Google Scholar]
  11. Cassimeris L., McNeill H., Zigmond S. H. Chemoattractant-stimulated polymorphonuclear leukocytes contain two populations of actin filaments that differ in their spatial distributions and relative stabilities. J Cell Biol. 1990 Apr;110(4):1067–1075. doi: 10.1083/jcb.110.4.1067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cockcroft S., Howell T. W., Gomperts B. D. Two G-proteins act in series to control stimulus-secretion coupling in mast cells: use of neomycin to distinguish between G-proteins controlling polyphosphoinositide phosphodiesterase and exocytosis. J Cell Biol. 1987 Dec;105(6 Pt 1):2745–2750. doi: 10.1083/jcb.105.6.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cooper J. A. Effects of cytochalasin and phalloidin on actin. J Cell Biol. 1987 Oct;105(4):1473–1478. doi: 10.1083/jcb.105.4.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Downey G. P., Chan C. K., Lea P., Takai A., Grinstein S. Phorbol ester-induced actin assembly in neutrophils: role of protein kinase C. J Cell Biol. 1992 Feb;116(3):695–706. doi: 10.1083/jcb.116.3.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Downey G. P., Grinstein S. Receptor-mediated actin assembly in electropermeabilized neutrophils: role of intracellular pH. Biochem Biophys Res Commun. 1989 Apr 14;160(1):18–24. doi: 10.1016/0006-291x(89)91614-8. [DOI] [PubMed] [Google Scholar]
  16. Downward J. The ras superfamily of small GTP-binding proteins. Trends Biochem Sci. 1990 Dec;15(12):469–472. doi: 10.1016/0968-0004(90)90300-z. [DOI] [PubMed] [Google Scholar]
  17. Egan S. E., Giddings B. W., Brooks M. W., Buday L., Sizeland A. M., Weinberg R. A. Association of Sos Ras exchange protein with Grb2 is implicated in tyrosine kinase signal transduction and transformation. Nature. 1993 May 6;363(6424):45–51. doi: 10.1038/363045a0. [DOI] [PubMed] [Google Scholar]
  18. Faulstich H., Zobeley S., Rinnerthaler G., Small J. V. Fluorescent phallotoxins as probes for filamentous actin. J Muscle Res Cell Motil. 1988 Oct;9(5):370–383. doi: 10.1007/BF01774064. [DOI] [PubMed] [Google Scholar]
  19. Hall A. Ras-related GTPases and the cytoskeleton. Mol Biol Cell. 1992 May;3(5):475–479. doi: 10.1091/mbc.3.5.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hall A. Ras-related proteins. Curr Opin Cell Biol. 1993 Apr;5(2):265–268. doi: 10.1016/0955-0674(93)90114-6. [DOI] [PubMed] [Google Scholar]
  21. Heath J. P. Behaviour and structure of the leading lamella in moving fibroblasts. I. Occurrence and centripetal movement of arc-shaped microfilament bundles beneath the dorsal cell surface. J Cell Sci. 1983 Mar;60:331–354. doi: 10.1242/jcs.60.1.331. [DOI] [PubMed] [Google Scholar]
  22. Herman I. M. Actin isoforms. Curr Opin Cell Biol. 1993 Feb;5(1):48–55. doi: 10.1016/s0955-0674(05)80007-9. [DOI] [PubMed] [Google Scholar]
  23. Hiraoka K., Kaibuchi K., Ando S., Musha T., Takaishi K., Mizuno T., Asada M., Ménard L., Tomhave E., Didsbury J. Both stimulatory and inhibitory GDP/GTP exchange proteins, smg GDS and rho GDI, are active on multiple small GTP-binding proteins. Biochem Biophys Res Commun. 1992 Jan 31;182(2):921–930. doi: 10.1016/0006-291x(92)91820-g. [DOI] [PubMed] [Google Scholar]
  24. Kahn R. A. Fluoride is not an activator of the smaller (20-25 kDa) GTP-binding proteins. J Biol Chem. 1991 Aug 25;266(24):15595–15597. [PubMed] [Google Scholar]
  25. Kawamoto S., Hidaka H. 1-(5-Isoquinolinesulfonyl)-2-methylpiperazine (H-7) is a selective inhibitor of protein kinase C in rabbit platelets. Biochem Biophys Res Commun. 1984 Nov 30;125(1):258–264. doi: 10.1016/s0006-291x(84)80362-9. [DOI] [PubMed] [Google Scholar]
  26. Koffer A., Edgar A. J., Bamburg J. R. Identification of two species of actin depolymerizing factor in cultures of BHK cells. J Muscle Res Cell Motil. 1988 Aug;9(4):320–328. doi: 10.1007/BF01773875. [DOI] [PubMed] [Google Scholar]
  27. Koffer A., Gomperts B. D. Soluble proteins as modulators of the exocytotic reaction of permeabilised rat mast cells. J Cell Sci. 1989 Nov;94(Pt 3):585–591. doi: 10.1242/jcs.94.3.585. [DOI] [PubMed] [Google Scholar]
  28. Koffer A., Tatham P. E., Gomperts B. D. Changes in the state of actin during the exocytotic reaction of permeabilized rat mast cells. J Cell Biol. 1990 Sep;111(3):919–927. doi: 10.1083/jcb.111.3.919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  30. Lassing I., Lindberg U. Evidence that the phosphatidylinositol cycle is linked to cell motility. Exp Cell Res. 1988 Jan;174(1):1–15. doi: 10.1016/0014-4827(88)90136-x. [DOI] [PubMed] [Google Scholar]
  31. Lillie T. H., Gomperts B. D. Kinetic characterization of guanine-nucleotide-induced exocytosis from permeabilized rat mast cells. Biochem J. 1993 Mar 1;290(Pt 2):389–394. doi: 10.1042/bj2900389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Maciver S. K., Zot H. G., Pollard T. D. Characterization of actin filament severing by actophorin from Acanthamoeba castellanii. J Cell Biol. 1991 Dec;115(6):1611–1620. doi: 10.1083/jcb.115.6.1611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Mousli M., Bronner C., Landry Y., Bockaert J., Rouot B. Direct activation of GTP-binding regulatory proteins (G-proteins) by substance P and compound 48/80. FEBS Lett. 1990 Jan 1;259(2):260–262. doi: 10.1016/0014-5793(90)80023-c. [DOI] [PubMed] [Google Scholar]
  34. Oberhauser A. F., Monck J. R., Balch W. E., Fernandez J. M. Exocytotic fusion is activated by Rab3a peptides. Nature. 1992 Nov 19;360(6401):270–273. doi: 10.1038/360270a0. [DOI] [PubMed] [Google Scholar]
  35. Philips M. R., Abramson S. B., Kolasinski S. L., Haines K. A., Weissmann G., Rosenfeld M. G. Low molecular weight GTP-binding proteins in human neutrophil granule membranes. J Biol Chem. 1991 Jan 15;266(2):1289–1298. [PubMed] [Google Scholar]
  36. Pollard T. D., Cooper J. A. Actin and actin-binding proteins. A critical evaluation of mechanisms and functions. Annu Rev Biochem. 1986;55:987–1035. doi: 10.1146/annurev.bi.55.070186.005011. [DOI] [PubMed] [Google Scholar]
  37. Quinn M. T., Evans T., Loetterle L. R., Jesaitis A. J., Bokoch G. M. Translocation of Rac correlates with NADPH oxidase activation. Evidence for equimolar translocation of oxidase components. J Biol Chem. 1993 Oct 5;268(28):20983–20987. [PubMed] [Google Scholar]
  38. Ridley A. J., Hall A. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell. 1992 Aug 7;70(3):389–399. doi: 10.1016/0092-8674(92)90163-7. [DOI] [PubMed] [Google Scholar]
  39. Ridley A. J., Paterson H. F., Johnston C. L., Diekmann D., Hall A. The small GTP-binding protein rac regulates growth factor-induced membrane ruffling. Cell. 1992 Aug 7;70(3):401–410. doi: 10.1016/0092-8674(92)90164-8. [DOI] [PubMed] [Google Scholar]
  40. Rodríguez Del Castillo A., Vitale M. L., Trifaró J. M. Ca2+ and pH determine the interaction of chromaffin cell scinderin with phosphatidylserine and phosphatidylinositol 4,5,-biphosphate and its cellular distribution during nicotinic-receptor stimulation and protein kinase C activation. J Cell Biol. 1992 Nov;119(4):797–810. doi: 10.1083/jcb.119.4.797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sontag J. M., Aunis D., Bader M. F. Peripheral actin filaments control calcium-mediated catecholamine release from streptolysin-O-permeabilized chromaffin cells. Eur J Cell Biol. 1988 Jun;46(2):316–326. [PubMed] [Google Scholar]
  42. Stossel T. P. From signal to pseudopod. How cells control cytoplasmic actin assembly. J Biol Chem. 1989 Nov 5;264(31):18261–18264. [PubMed] [Google Scholar]
  43. Takai Y., Kaibuchi K., Kikuchi A., Kawata M. Small GTP-binding proteins. Int Rev Cytol. 1992;133:187–230. doi: 10.1016/s0074-7696(08)61861-6. [DOI] [PubMed] [Google Scholar]
  44. Theriot J. A., Mitchison T. J. The nucleation-release model of actin filament dynamics in cell motility. Trends Cell Biol. 1992 Aug;2(8):219–222. doi: 10.1016/0962-8924(92)90298-2. [DOI] [PubMed] [Google Scholar]
  45. Therrien S., Naccache P. H. Guanine nucleotide-induced polymerization of actin in electropermeabilized human neutrophils. J Cell Biol. 1989 Sep;109(3):1125–1132. doi: 10.1083/jcb.109.3.1125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Yonezawa N., Nishida E., Iida K., Yahara I., Sakai H. Inhibition of the interactions of cofilin, destrin, and deoxyribonuclease I with actin by phosphoinositides. J Biol Chem. 1990 May 25;265(15):8382–8386. [PubMed] [Google Scholar]
  47. Zhang J., King W. G., Dillon S., Hall A., Feig L., Rittenhouse S. E. Activation of platelet phosphatidylinositide 3-kinase requires the small GTP-binding protein Rho. J Biol Chem. 1993 Oct 25;268(30):22251–22254. [PubMed] [Google Scholar]
  48. de Hostos E. L., Bradtke B., Lottspeich F., Guggenheim R., Gerisch G. Coronin, an actin binding protein of Dictyostelium discoideum localized to cell surface projections, has sequence similarities to G protein beta subunits. EMBO J. 1991 Dec;10(13):4097–4104. doi: 10.1002/j.1460-2075.1991.tb04986.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES