Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1991 Apr 2;113(2):321–330. doi: 10.1083/jcb.113.2.321

Radixin, a barbed end-capping actin-modulating protein, is concentrated at the cleavage furrow during cytokinesis [published erratum appears in J Cell Biol 1991 Sep;114(5):1101-3]

PMCID: PMC2288944  PMID: 1707055

Abstract

Radixin is a barbed end-capping actin-modulating protein which was first identified in isolated cell-to-cell adherens junctions from rat liver (Tsukita, Sa., Y. Hieda, and Sh. Tsukita, 1989. J. Cell Biol. 108:2369-2382). In the present study, we have analyzed the distribution of radixin in dividing cells. For this purpose, an mAb specific for radixin was obtained using chicken gizzard radixin as an antigen. By immunofluorescence microscopy with this mAb and a polyclonal antibody obtained previously, it was clearly shown in rat fibroblastic cells (3Y1 cells) that radixin was highly concentrated at the cleavage furrow during cytokinesis. Radixin appeared to accumulate rapidly at the cleavage furrow at the onset of furrowing, continued to be concentrated at the furrow during anaphase and telophase, and was finally enriched at the midbody. This concentration of radixin at the cleavage furrow was detected in all other cultured cells we examined: bovine epithelial cells (MDBK cells), mouse myeloma cells (P3 cells), rat kangaroo Ptk2 cells, mouse teratocarcinoma cells, and chicken fibroblasts. Furthermore, it became clear that the epitope for the mAb was immunofluorescently masked in the cell-to-cell adherens junctions. Together, these results lead us to conclude that radixin is present in the undercoat of the cell-to-cell adherens junctions and that of the cleavage furrow, although their respective molecular architectures are distinct. The possible roles of radixin at the cleavage furrow are discussed with special reference to the molecular mechanism of the actin filament-plasma membrane interaction at the furrow.

Full Text

The Full Text of this article is available as a PDF (3.2 MB).

Selected References

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

  1. Aubin J. E., Weber K., Osborn M. Analysis of actin and microfilament-associated proteins in the mitotic spindle and cleavage furrow of PtK2 cells by immunofluorescence microscopy. A critical note. Exp Cell Res. 1979 Nov;124(1):93–109. doi: 10.1016/0014-4827(79)90260-x. [DOI] [PubMed] [Google Scholar]
  2. Burridge K., Connell L. A new protein of adhesion plaques and ruffling membranes. J Cell Biol. 1983 Aug;97(2):359–367. doi: 10.1083/jcb.97.2.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burridge K., Mangeat P. An interaction between vinculin and talin. Nature. 1984 Apr 19;308(5961):744–746. doi: 10.1038/308744a0. [DOI] [PubMed] [Google Scholar]
  4. Byers H. R., White G. E., Fujiwara K. Organization and function of stress fibers in cells in vitro and in situ. A review. Cell Muscle Motil. 1984;5:83–137. doi: 10.1007/978-1-4684-4592-3_2. [DOI] [PubMed] [Google Scholar]
  5. Cao L. G., Wang Y. L. Mechanism of the formation of contractile ring in dividing cultured animal cells. I. Recruitment of preexisting actin filaments into the cleavage furrow. J Cell Biol. 1990 Apr;110(4):1089–1095. doi: 10.1083/jcb.110.4.1089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  7. Cowin P., Kapprell H. P., Franke W. W., Tamkun J., Hynes R. O. Plakoglobin: a protein common to different kinds of intercellular adhering junctions. Cell. 1986 Sep 26;46(7):1063–1073. doi: 10.1016/0092-8674(86)90706-3. [DOI] [PubMed] [Google Scholar]
  8. De Blas A. L., Cherwinski H. M. Detection of antigens on nitrocellulose paper immunoblots with monoclonal antibodies. Anal Biochem. 1983 Aug;133(1):214–219. doi: 10.1016/0003-2697(83)90245-2. [DOI] [PubMed] [Google Scholar]
  9. FARQUHAR M. G., PALADE G. E. Junctional complexes in various epithelia. J Cell Biol. 1963 May;17:375–412. doi: 10.1083/jcb.17.2.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Feramisco J. R., Burridge K. A rapid purification of alpha-actinin, filamin, and a 130,000-dalton protein from smooth muscle. J Biol Chem. 1980 Feb 10;255(3):1194–1199. [PubMed] [Google Scholar]
  11. Fujiwara K., Porter M. E., Pollard T. D. Alpha-actinin localization in the cleavage furrow during cytokinesis. J Cell Biol. 1978 Oct;79(1):268–275. doi: 10.1083/jcb.79.1.268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Geiger B. A 130K protein from chicken gizzard: its localization at the termini of microfilament bundles in cultured chicken cells. Cell. 1979 Sep;18(1):193–205. doi: 10.1016/0092-8674(79)90368-4. [DOI] [PubMed] [Google Scholar]
  13. Geiger B., Avnur Z., Rinnerthaler G., Hinssen H., Small V. J. Microfilament-organizing centers in areas of cell contact: cytoskeletal interactions during cell attachment and locomotion. J Cell Biol. 1984 Jul;99(1 Pt 2):83s–91s. doi: 10.1083/jcb.99.1.83s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Geiger B. Membrane-cytoskeleton interaction. Biochim Biophys Acta. 1983 Aug 11;737(3-4):305–341. doi: 10.1016/0304-4157(83)90005-9. [DOI] [PubMed] [Google Scholar]
  15. Geiger B., Tokuyasu K. T., Dutton A. H., Singer S. J. Vinculin, an intracellular protein localized at specialized sites where microfilament bundles terminate at cell membranes. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4127–4131. doi: 10.1073/pnas.77.7.4127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Geiger B., Volk T., Volberg T. Molecular heterogeneity of adherens junctions. J Cell Biol. 1985 Oct;101(4):1523–1531. doi: 10.1083/jcb.101.4.1523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Herman I. M., Pollard T. D. Comparison of purified anti-actin and fluorescent-heavy meromyosin staining patterns in dividing cells. J Cell Biol. 1979 Mar;80(3):509–520. doi: 10.1083/jcb.80.3.509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Lazarides E., Burridge K. Alpha-actinin: immunofluorescent localization of a muscle structural protein in nonmuscle cells. Cell. 1975 Nov;6(3):289–298. doi: 10.1016/0092-8674(75)90180-4. [DOI] [PubMed] [Google Scholar]
  20. Mabuchi I. Biochemical aspects of cytokinesis. Int Rev Cytol. 1986;101:175–213. doi: 10.1016/s0074-7696(08)60249-1. [DOI] [PubMed] [Google Scholar]
  21. Mabuchi I., Hamaguchi Y., Kobayashi T., Hosoya H., Tsukita S., Tsukita S. Alpha-actinin from sea urchin eggs: biochemical properties, interaction with actin, and distribution in the cell during fertilization and cleavage. J Cell Biol. 1985 Feb;100(2):375–383. doi: 10.1083/jcb.100.2.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mabuchi I., Tsukita S., Tsukita S., Sawai T. Cleavage furrow isolated from newt eggs: contraction, organization of the actin filaments, and protein components of the furrow. Proc Natl Acad Sci U S A. 1988 Aug;85(16):5966–5970. doi: 10.1073/pnas.85.16.5966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nunnally M. H., D'Angelo J. M., Craig S. W. Filamin concentration in cleavage furrow and midbody region: frequency of occurrence compared with that of alpha-actinin and myosin. J Cell Biol. 1980 Oct;87(1):219–226. doi: 10.1083/jcb.87.1.219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ohba H., Ohmori I., Numata O., Watanabe Y. Purification and immunofluorescence localization of the mutant gene product of a Tetrahymena cdaA1 mutant affecting cell division. J Biochem. 1986 Sep;100(3):797–808. doi: 10.1093/oxfordjournals.jbchem.a121773. [DOI] [PubMed] [Google Scholar]
  25. Perry M. M., John H. A., Thomas N. S. Actin-like filaments in the cleavage furrow of newt egg. Exp Cell Res. 1971 Mar;65(1):249–253. doi: 10.1016/s0014-4827(71)80075-7. [DOI] [PubMed] [Google Scholar]
  26. Pollard T. D., Weihing R. R. Actin and myosin and cell movement. CRC Crit Rev Biochem. 1974 Jan;2(1):1–65. doi: 10.3109/10409237409105443. [DOI] [PubMed] [Google Scholar]
  27. Rappaport R. Establishment of the mechanism of cytokinesis in animal cells. Int Rev Cytol. 1986;105:245–281. doi: 10.1016/s0074-7696(08)61065-7. [DOI] [PubMed] [Google Scholar]
  28. Sanger J. M., Mittal B., Pochapin M. B., Sanger J. W. Stress fiber and cleavage furrow formation in living cells microinjected with fluorescently labeled alpha-actinin. Cell Motil Cytoskeleton. 1987;7(3):209–220. doi: 10.1002/cm.970070304. [DOI] [PubMed] [Google Scholar]
  29. Sanger J. W. Changing patterns of actin localization during cell division. Proc Natl Acad Sci U S A. 1975 May;72(5):1913–1916. doi: 10.1073/pnas.72.5.1913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schroeder T. E. Actin in dividing cells: contractile ring filaments bind heavy meromyosin. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1688–1692. doi: 10.1073/pnas.70.6.1688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schroeder T. E. The contractile ring. I. Fine structure of dividing mammalian (HeLa) cells and the effects of cytochalasin B. Z Zellforsch Mikrosk Anat. 1970;109(4):431–449. [PubMed] [Google Scholar]
  32. Schroeder T. E. The contractile ring. II. Determining its brief existence, volumetric changes, and vital role in cleaving Arbacia eggs. J Cell Biol. 1972 May;53(2):419–434. doi: 10.1083/jcb.53.2.419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Staehelin L. A. Structure and function of intercellular junctions. Int Rev Cytol. 1974;39:191–283. doi: 10.1016/s0074-7696(08)60940-7. [DOI] [PubMed] [Google Scholar]
  34. Tsukita S., Hieda Y., Tsukita S. A new 82-kD barbed end-capping protein (radixin) localized in the cell-to-cell adherens junction: purification and characterization. J Cell Biol. 1989 Jun;108(6):2369–2382. doi: 10.1083/jcb.108.6.2369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tsukita S., Itoh M., Tsukita S. A new 400-kD protein from isolated adherens junctions: its localization at the undercoat of adherens junctions and at microfilament bundles such as stress fibers and circumferential bundles. J Cell Biol. 1989 Dec;109(6 Pt 1):2905–2915. doi: 10.1083/jcb.109.6.2905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tsukita S., Tsukita S. Isolation of cell-to-cell adherens junctions from rat liver. J Cell Biol. 1989 Jan;108(1):31–41. doi: 10.1083/jcb.108.1.31. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tsukita S., Tsukita S., Nagafuchi A. The undercoat of adherens junctions: a key specialized structure in organogenesis and carcinogenesis. Cell Struct Funct. 1990 Feb;15(1):7–12. doi: 10.1247/csf.15.7. [DOI] [PubMed] [Google Scholar]
  38. Vaessen R. T., Kreike J., Groot G. S. Protein transfer to nitrocellulose filters. A simple method for quantitation of single proteins in complex mixtures. FEBS Lett. 1981 Feb 23;124(2):193–196. doi: 10.1016/0014-5793(81)80134-2. [DOI] [PubMed] [Google Scholar]
  39. Volberg T., Geiger B., Kartenbeck J., Franke W. W. Changes in membrane-microfilament interaction in intercellular adherens junctions upon removal of extracellular Ca2+ ions. J Cell Biol. 1986 May;102(5):1832–1842. doi: 10.1083/jcb.102.5.1832. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wang Y. L. Mobility of filamentous actin in living cytoplasm. J Cell Biol. 1987 Dec;105(6 Pt 1):2811–2816. doi: 10.1083/jcb.105.6.2811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wessells N. K., Spooner B. S., Ash J. F., Bradley M. O., Luduena M. A., Taylor E. L., Wrenn J. T., Yamada K. Microfilaments in cellular and developmental processes. Science. 1971 Jan 15;171(3967):135–143. doi: 10.1126/science.171.3967.135. [DOI] [PubMed] [Google Scholar]

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

RESOURCES