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. 1976 Dec 1;71(3):704–714. doi: 10.1083/jcb.71.3.704

Actin polymerization and interaction with other proteins in temperature- induced gelation of sea urchin egg extracts

PMCID: PMC2109792  PMID: 1033188

Abstract

The gel which forms on warming the extracts of the cytoplasmic proteins of sea urchin eggs has been separated into two fractions, one containing F-actin and the other containing two proteins of 58,000 and 22,000 mol wt. When combined in 0.1 M KCl, even at 0 degrees C, these components will form gel material identical to that formed by warming extracts. This gel is a network of laterally aggregated F-actin filaments which are in register and which display a complex cross- banding pattern generated by the presence of the other two proteins. Low concentrations of calcium block the assembly of these proteins to form this complex structure, which may play some cytoskeletal role in the cytoplasm. This association of F-actin with the other proteins to form a gel is very likely the last step fo the process occurring in warmed extracts. At low temperatures, gelation of extracts is limited by the relative absence of F-actin, as demonstrated by the inability to sediment it at 100,000 g and also by the fact that gelation occurs immediately if exogenous F-actin is added to cold extracts. The transformation of the G-actin present in the extract to the F-form is apparently repressed at low temperatures. This is shown directly by the failure of added G-actin to polymerize at low temperatures in the presence of extract. These observations resemble those which have been reported on preparations from amoeboid cells and may be significant in the involvement of actin and these other proteins in cell division and later developmental processes.

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Selected References

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  1. Bray D., Thomas C. The actin content of fibroblasts. Biochem J. 1975 May;147(2):221–228. doi: 10.1042/bj1470221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Gillis J. M., O'Brien E. J. The effect of calcium ions on the structure of reconstituted muscle thin filaments. J Mol Biol. 1975 Dec 15;99(3):445–459. doi: 10.1016/s0022-2836(75)80137-9. [DOI] [PubMed] [Google Scholar]
  3. Gustafson T., Wolpert L. Cellular movement and contact in sea urchin morphogenesis. Biol Rev Camb Philos Soc. 1967 Aug;42(3):442–498. doi: 10.1111/j.1469-185x.1967.tb01482.x. [DOI] [PubMed] [Google Scholar]
  4. Hanson J. Evidence from electron microscope studies on actin paracrystals concerning the origin of the cross-striation in the thin filaments of vertebrate skeletal muscle. Proc R Soc Lond B Biol Sci. 1973 Feb 27;183(1070):39–58. doi: 10.1098/rspb.1973.0003. [DOI] [PubMed] [Google Scholar]
  5. Hartwig J. H., Stossel T. P. Isolation and properties of actin, myosin, and a new actinbinding protein in rabbit alveolar macrophages. J Biol Chem. 1975 Jul 25;250(14):5696–5705. [PubMed] [Google Scholar]
  6. Kane R. E. Preparation and purification of polymerized actin from sea urchin egg extracts. J Cell Biol. 1975 Aug;66(2):305–315. doi: 10.1083/jcb.66.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  8. Morgan J. Microfilaments from amoeba proteins. Exp Cell Res. 1971 Mar;65(1):7–16. doi: 10.1016/s0014-4827(71)80043-5. [DOI] [PubMed] [Google Scholar]
  9. O'Brien E. J., Gillis J. M., Couch J. Symmetry and molecular arrangement in paracrystals of reconstituted muscle thin filaments. J Mol Biol. 1975 Dec 15;99(3):461–475. doi: 10.1016/s0022-2836(75)80138-0. [DOI] [PubMed] [Google Scholar]
  10. Pollard T. D., Ito S. Cytoplasmic filaments of Amoeba proteus. I. The role of filaments in consistency changes and movement. J Cell Biol. 1970 Aug;46(2):267–289. doi: 10.1083/jcb.46.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Pollard T. D., Korn E. D. Filaments of Amoeba proteus. II. Binding of heavy meromyosin by thin filaments in motile cytoplasmic extracts. J Cell Biol. 1971 Jan;48(1):216–219. doi: 10.1083/jcb.48.1.216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. 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]
  15. Spudich J. A., Cooke R. Supramolecular forms of actin from amoebae of Dictyostelium discoideum. J Biol Chem. 1975 Sep 25;250(18):7485–7491. [PubMed] [Google Scholar]
  16. Spudich J. A., Huxley H. E., Finch J. T. Regulation of skeletal muscle contraction. II. Structural studies of the interaction of the tropomyosin-troponin complex with actin. J Mol Biol. 1972 Dec 30;72(3):619–632. doi: 10.1016/0022-2836(72)90180-5. [DOI] [PubMed] [Google Scholar]
  17. Stossel T. P., Hartwig J. H. Interactions of actin, myosin, and a new actin-binding protein of rabbit pulmonary macrophages. II. Role in cytoplasmic movement and phagocytosis. J Cell Biol. 1976 Mar;68(3):602–619. doi: 10.1083/jcb.68.3.602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. THOMPSON C. M., WOLPERT L. THE ISOLATION OF MOTILE CYTOPLASM FROM AMOEBA PROTEUS. Exp Cell Res. 1963 Oct;32:156–160. doi: 10.1016/0014-4827(63)90078-8. [DOI] [PubMed] [Google Scholar]
  19. Taylor D. L., Condeelis J. S., Moore P. L., Allen R. D. The contractile basis of amoeboid movement. I. The chemical control of motility in isolated cytoplasm. J Cell Biol. 1973 Nov;59(2 Pt 1):378–394. doi: 10.1083/jcb.59.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tilney L. G. Actin filaments in the acrosomal reaction of Limulus sperm. Motion generated by alterations in the packing of the filaments. J Cell Biol. 1975 Feb;64(2):289–310. doi: 10.1083/jcb.64.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wakabayashi T., Huxley H. E., Amos L. A., Klug A. Three-dimensional image reconstruction of actin-tropomyosin complex and actin-tropomyosin-troponin T-troponin I complex. J Mol Biol. 1975 Apr 25;93(4):477–497. doi: 10.1016/0022-2836(75)90241-7. [DOI] [PubMed] [Google Scholar]
  22. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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