Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1982 Apr 1;93(1):122–128. doi: 10.1083/jcb.93.1.122

On the dynamics of the microfilament system in HeLa cells

I Blikstad, L Carlsson
PMCID: PMC2112103  PMID: 6802855

Abstract

We measured the pools of unpolymerized and filamentous actin in homogenates of HeLa cells made in several different lysis buffers, as well as after treatment of cells with a variety of chemicals or trypsin, and after adenovirus (type 2) infection. This was possible when a series of factors concerning the basic culture conditions were kept constant: e.g., serum type used, serum batch, cell density, time after subcultivation of cells, and buffering substance in the medium. Homogenates from untreated cells usually contain 35-45 percent of the total actin in an unpolymerized form. With some batches of cells this number can be as high as 50 percent. In sparse cultures (3 x 10(4) cell/cm(2)), HeLa cells contain approximately 10 pg actin/cell, while the corresponding number is only 5 pg in dense cultures (3 x 10(5) cells/cm(2)). Treatment of cells with cytochalasin B increases the pool of unpolymerized actin by approximately 30-40 percent, while colchicine decreases the fraction of unpolymerized actin by 20 percent. The oxidant diamide increases the filamentous actin pool 25-50 percent. Glucose, sodium azide, dinitrophenol, serum starvation, or thymidine treatment does not affect the distribution between unpolymerized and filamentous actin to any significant extent. Trypsin and EDTA induced rounding up of cells but did not change the actin distribution. The distribution of actin between G- and F-forms was unchanged after adenovirus infection. These results show that significant changes in the actin pools can be induced in nucleated cells. However, several treatments which alter the morphology and motility of cells are not accompanied by an alteration in the G-/F-actin ratio.

Full Text

The Full Text of this article is available as a PDF (807.1 KB).

Selected References

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

  1. Blikstad I., Markey F., Carlsson L., Persson T., Lindberg U. Selective assay of monomeric and filamentous actin in cell extracts, using inhibition of deoxyribonuclease I. Cell. 1978 Nov;15(3):935–943. doi: 10.1016/0092-8674(78)90277-5. [DOI] [PubMed] [Google Scholar]
  2. Brenner S. L., Korn E. D. Substoichiometric concentrations of cytochalasin D inhibit actin polymerization. Additional evidence for an F-actin treadmill. J Biol Chem. 1979 Oct 25;254(20):9982–9985. [PubMed] [Google Scholar]
  3. Brown S. S., Spudich J. A. Cytochalasin inhibits the rate of elongation of actin filament fragments. J Cell Biol. 1979 Dec;83(3):657–662. doi: 10.1083/jcb.83.3.657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carlsson L., Blikstad I. Colchicine treatment of HeLa cells alters the G/F actin ratio. FEBS Lett. 1981 Feb 23;124(2):282–284. doi: 10.1016/0014-5793(81)80156-1. [DOI] [PubMed] [Google Scholar]
  5. Carlsson L., Markey F., Blikstad I., Persson T., Lindberg U. Reorganization of actin in platelets stimulated by thrombin as measured by the DNase I inhibition assay. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6376–6380. doi: 10.1073/pnas.76.12.6376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carlsson L., Nyström L. E., Sundkvist I., Markey F., Lindberg U. Actin polymerizability is influenced by profilin, a low molecular weight protein in non-muscle cells. J Mol Biol. 1977 Sep 25;115(3):465–483. doi: 10.1016/0022-2836(77)90166-8. [DOI] [PubMed] [Google Scholar]
  7. Franke W. W., Schmid E., Weber K., Osborn M. HeLa cells contain intermediate-sized filaments of the prekeratin type. Exp Cell Res. 1979 Jan;118(1):95–109. doi: 10.1016/0014-4827(79)90587-1. [DOI] [PubMed] [Google Scholar]
  8. Goldman R. D. The role of three cytoplasmic fibers in BHK-21 cell motility. I. Microtubules and the effects of colchicine. J Cell Biol. 1971 Dec;51(3):752–762. doi: 10.1083/jcb.51.3.752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hartwig J. H., Stossel T. P. Interactions of actin, myosin, and an actin-binding protein of rabbit pulmonary macrophages. III. Effects of cytochalasin B. J Cell Biol. 1976 Oct;71(1):295–303. doi: 10.1083/jcb.71.1.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Laub F., Kaplan M., Gitler C. Actin polymerization accompanies Thy-1-capping on mouse thymocytes. FEBS Lett. 1981 Feb 9;124(1):35–38. doi: 10.1016/0014-5793(81)80048-8. [DOI] [PubMed] [Google Scholar]
  11. Lazarides E. Intermediate filaments as mechanical integrators of cellular space. Nature. 1980 Jan 17;283(5744):249–256. doi: 10.1038/283249a0. [DOI] [PubMed] [Google Scholar]
  12. Lin D. C., Lin S. Actin polymerization induced by a motility-related high-affinity cytochalasin binding complex from human erythrocyte membrane. Proc Natl Acad Sci U S A. 1979 May;76(5):2345–2349. doi: 10.1073/pnas.76.5.2345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lin D. C., Tobin K. D., Grumet M., Lin S. Cytochalasins inhibit nuclei-induced actin polymerization by blocking filament elongation. J Cell Biol. 1980 Feb;84(2):455–460. doi: 10.1083/jcb.84.2.455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. MacLean-Fletcher S., Pollard T. D. Mechanism of action of cytochalasin B on actin. Cell. 1980 Jun;20(2):329–341. doi: 10.1016/0092-8674(80)90619-4. [DOI] [PubMed] [Google Scholar]
  15. Morris A., Tannenbaum J. Cytochalasin D does not produce net depolymerization of actin filaments in HEp-2 cells. Nature. 1980 Oct 16;287(5783):637–639. doi: 10.1038/287637a0. [DOI] [PubMed] [Google Scholar]
  16. Nath J., Rebhun J. I. Effects of caffeine and other methylxanthines on the development and metabolism of sea urchin eggs. Involvement of NADP and glutathione. J Cell Biol. 1976 Mar;68(3):440–450. doi: 10.1083/jcb.68.3.440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Oliver J. M., Albertini D. F., Berlin R. D. Effects of glutathione-oxidizing agents on microtubule assembly and microtubule-dependent surface properties of human neutrophils. J Cell Biol. 1976 Dec;71(3):921–932. doi: 10.1083/jcb.71.3.921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Osborn M., Weber K. Cytoplasmic microtubules in tissue culture cells appear to grow from an organizing structure towards the plasma membrane. Proc Natl Acad Sci U S A. 1976 Mar;73(3):867–871. doi: 10.1073/pnas.73.3.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ostlund R. E., Jr, Leung J. T., Hajek S. V. Regulation of microtubule assembly in cultured fibroblasts. J Cell Biol. 1980 May;85(2):386–391. doi: 10.1083/jcb.85.2.386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Otto J. J., Kane R. E., Bryan J. Redistribution of actin and fascin in sea urchin eggs after fertilization. Cell Motil. 1980;1(1):31–40. doi: 10.1002/cm.970010104. [DOI] [PubMed] [Google Scholar]
  21. Pollard T. D. Cytoskeletal functions of cytoplasmic contractile proteins. J Supramol Struct. 1976;5(3):317–334. doi: 10.1002/jss.400050306. [DOI] [PubMed] [Google Scholar]
  22. Swanston-Flatt S. K., Carlsson L., Gylfe E. Actin filament formation in pancreatic beta-cells during glucose stimulation of insulin secretion. FEBS Lett. 1980 Aug 11;117(1):299–302. doi: 10.1016/0014-5793(80)80966-5. [DOI] [PubMed] [Google Scholar]
  23. Tilney L. G. Studies on the microtubules in heliozoa. IV. The effect of colchicine on the formation and maintenance of the axopodia and the redevelopment of pattern in Actinosphaerium nucleofilum (Barrett). J Cell Sci. 1968 Dec;3(4):549–562. doi: 10.1242/jcs.3.4.549. [DOI] [PubMed] [Google Scholar]
  24. Weihing R. R. Cytochalasin B inhibits actin-related gelation of HeLa cell extracts. J Cell Biol. 1976 Oct;71(1):303–307. doi: 10.1083/jcb.71.1.303. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES