Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1988 Oct 1;107(4):1499–1504. doi: 10.1083/jcb.107.4.1499

Functional alterations in beta'-actin from a KB cell mutant resistant to cytochalasin B

PMCID: PMC2115235  PMID: 3049623

Abstract

We recently described the isolation of mutant KB cells (Cyt 1 cells) resistant to the cytotoxic effect of cytochalasin B (CB). This mutant carried an altered beta-actin; i.e., beta'-actin (Toyama, S., and S. Toyama. 1984. Cell. 37:609-614). In the present study, we have examined the functional properties of actin in Cyt 1 cells. Our results showed that increased resistance of Cyt 1 cells to CB was reflected in altered properties of beta'-actin itself. This was shown directly by two findings. First, the polymerization of beta'-actin was more resistant than that of beta- or gamma-actin to the multiple effects of CB. Second, beta'-actin bound less CB than beta- or gamma-actin. The functional alteration of beta'-actin in Cyt 1 cells was further supported by the observation that, although treatment of KB cells with CB increased the pool of unpolymerized actin, the same treatment did not affect the pool of unpolymerized actin in Cyt 1 cells, and that microfilaments of Cyt 1 cells were more resistant to the disrupting action of CB than those of KB cells. These results strongly suggest that the primary site of action of CB on cell motility processes is actin.

Full Text

The Full Text of this article is available as a PDF (1.5 MB).

Selected References

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

  1. Blikstad I., Carlsson L. On the dynamics of the microfilament system in HeLa cells. J Cell Biol. 1982 Apr;93(1):122–128. doi: 10.1083/jcb.93.1.122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Bonder E. M., Mooseker M. S. Cytochalasin B slows but does not prevent monomer addition at the barbed end of the actin filament. J Cell Biol. 1986 Jan;102(1):282–288. doi: 10.1083/jcb.102.1.282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Brown S. S., Spudich J. A. Mechanism of action of cytochalasin: evidence that it binds to actin filament ends. J Cell Biol. 1981 Mar;88(3):487–491. doi: 10.1083/jcb.88.3.487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Flanagan M. D., Lin S. Cytochalasins block actin filament elongation by binding to high affinity sites associated with F-actin. J Biol Chem. 1980 Feb 10;255(3):835–838. [PubMed] [Google Scholar]
  8. Goddette D. W., Frieden C. Actin polymerization. The mechanism of action of cytochalasin D. J Biol Chem. 1986 Dec 5;261(34):15974–15980. [PubMed] [Google Scholar]
  9. Goddette D. W., Frieden C. The binding of cytochalasin D to monomeric actin. Biochem Biophys Res Commun. 1985 May 16;128(3):1087–1092. doi: 10.1016/0006-291x(85)91051-4. [DOI] [PubMed] [Google Scholar]
  10. Gordon D. J., Eisenberg E., Korn E. D. Characterization of cytoplasmic actin isolated from Acanthamoeba castellanii by a new method. J Biol Chem. 1976 Aug 10;251(15):4778–4786. [PubMed] [Google Scholar]
  11. 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]
  12. Ishiura M., Okada Y. The role of actin in temperature-dependent gel-sol transformation of extracts of Ehrlich ascites tumor cells. J Cell Biol. 1979 Feb;80(2):465–480. doi: 10.1083/jcb.80.2.465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kouyama T., Mihashi K. Fluorimetry study of N-(1-pyrenyl)iodoacetamide-labelled F-actin. Local structural change of actin protomer both on polymerization and on binding of heavy meromyosin. Eur J Biochem. 1981;114(1):33–38. [PubMed] [Google Scholar]
  14. Lin D. C., Lin S. A rapid assay for actin-associated high-affinity cytochalasin binding sites based on isoelectric precipitation of soluble protein. Anal Biochem. 1980 Apr;103(2):316–322. doi: 10.1016/0003-2697(80)90617-x. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Löw I., Dancker P. Effect of cytochalasin B on formation and properties of muscle F-actin. Biochim Biophys Acta. 1976 May 14;430(2):366–374. doi: 10.1016/0005-2728(76)90092-x. [DOI] [PubMed] [Google Scholar]
  17. Mabuchi I. Electron microscopic determination of the actin filament end at which cytochalasin B blocks monomer addition using the acrosomal actin bundle from horseshoe crab sperm. J Biochem. 1983 Oct;94(4):1349–1352. doi: 10.1093/oxfordjournals.jbchem.a134480. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. McKenna N., Meigs J. B., Wang Y. L. Identical distribution of fluorescently labeled brain and muscle actins in living cardiac fibroblasts and myocytes. J Cell Biol. 1985 Jan;100(1):292–296. doi: 10.1083/jcb.100.1.292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Neefjes J. J., Breur-Vriesendorp B. S., van Seventer G. A., Iványi P., Ploegh H. L. An improved biochemical method for the analysis of HLA-class I antigens. Definition of new HLA-class I subtypes. Hum Immunol. 1986 Jun;16(2):169–181. doi: 10.1016/0198-8859(86)90046-7. [DOI] [PubMed] [Google Scholar]
  21. Northrop J., Weber A., Mooseker M. S., Franzini-Armstrong C., Bishop M. F., Dubyak G. R., Tucker M., Walsh T. P. Different calcium dependence of the capping and cutting activities of villin. J Biol Chem. 1986 Jul 15;261(20):9274–9281. [PubMed] [Google Scholar]
  22. Otey C. A., Kalnoski M. H., Lessard J. L., Bulinski J. C. Immunolocalization of the gamma isoform of nonmuscle actin in cultured cells. J Cell Biol. 1986 May;102(5):1726–1737. doi: 10.1083/jcb.102.5.1726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pollard T. D., Mooseker M. S. Direct measurement of actin polymerization rate constants by electron microscopy of actin filaments nucleated by isolated microvillus cores. J Cell Biol. 1981 Mar;88(3):654–659. doi: 10.1083/jcb.88.3.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pollard T. D. The role of actin in the temperature-dependent gelation and contraction of extracts of Acanthamoeba. J Cell Biol. 1976 Mar;68(3):579–601. doi: 10.1083/jcb.68.3.579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Segura M., Lindberg U. Separation of non-muscle isoactins in the free form or as profilactin complexes. J Biol Chem. 1984 Mar 25;259(6):3949–3954. [PubMed] [Google Scholar]
  26. Tellam R., Frieden C. Cytochalasin D and platelet gelsolin accelerate actin polymer formation. A model for regulation of the extent of actin polymer formation in vivo. Biochemistry. 1982 Jun 22;21(13):3207–3214. doi: 10.1021/bi00256a027. [DOI] [PubMed] [Google Scholar]
  27. Toyama S., Toyama S. A variant form of beta-actin in a mutant of KB cells resistant to cytochalasin B. Cell. 1984 Jun;37(2):609–614. doi: 10.1016/0092-8674(84)90391-x. [DOI] [PubMed] [Google Scholar]
  28. Toyama S., Toyama S., Uetake H. Altered cell-fusion capacity in lines of KB cells resistant to Sendai virus-induced cytolysis. Virology. 1977 Feb;76(2):503–515. doi: 10.1016/0042-6822(77)90233-1. [DOI] [PubMed] [Google Scholar]
  29. Weber K., Rathke P. C., Osborn M. Cytoplasmic microtubular images in glutaraldehyde-fixed tissue culture cells by electron microscopy and by immunofluorescence microscopy. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1820–1824. doi: 10.1073/pnas.75.4.1820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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