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PMID: 8603919 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Xenopus laevis actin-depolymerizing factor/cofilin: a phosphorylation-regulated protein essential for development.

The Journal of cell biology ·Vol. 132 ·No. 5 ·1996-03-00 ·Pages 871-85

Abe H, Obinata T, Minamide LS, Bamburg JR

Abstract

Two cDNAs, isolated from a Xenopus laevis embryonic library, encode proteins of 168 amino acids, both of which are 77% identical to chick cofilin and 66% identical to chick actin-depolymerizing factor (ADF), two structurally and functionally related proteins. These Xenopus ADF/cofilins (XADs) differ from each other in 12 residues spread throughout the sequence but do not differ in charge. Purified GST-fusion proteins have pH-dependent actin-depolymerizing and F-actin-binding activities similar to chick ADF and cofilin. Similarities in the developmental and tissue specific expression, embryonic localization, and in the cDNA sequence of the noncoding regions, suggest that the two XACs arise from allelic variants of the pseudotetraploid X. laevis. Immunofluorescence localization of XAC in oocyte sections with an XAC-specific monoclonal antibody shows it to be diffuse in the cortical cytoplasm. After fertilization, increased immunostaining is observed in two regions: along the membrane, particularly that of the vegetal hemisphere, and at the interface between the cortical and animal hemisphere cytoplasm. The cleavage furrow and the mid-body structure are stained at the end of first cleavage. Neuroectoderm derived tissues, notochord, somites, and epidermis stain heavily either continuously or transiently from stages 18-34. A phosphorylated form of XAC (pXAC) was identified by 2D Western blotting, and it is the only species found in oocytes. Dephosphorylation of >60% of the pXAC occurs within 30 min after fertilization. Injection of one blastomere at the 2 cell stage, either with constitutively active XAC or with an XAC inhibitory antibody, blocked cleavage of only the injected blastomere in a concentration-dependent manner without inhibiting nuclear division. The cleavage furrow of eggs injected with constitutively active XAC completely regressed. Blastomeres injected with neutralized antibody developed normally. These results suggest that XAC is necessary for cytokinesis and that its activity must be properly regulated for cleavage to occur.

MeSH Terms
Actin Depolymerizing Factors Alleles Amino Acid Sequence Animals Base Sequence Cell Compartmentation Cell Division/physiology Cofilin 1 Cofilin 2 Cytoskeletal Proteins DNA, Complementary/genetics Destrin Fluorescent Antibody Technique Gene Expression Regulation, Developmental Genes, Lethal In Situ Hybridization Microfilament Proteins/biosynthesis,genetics Molecular Sequence Data Nerve Tissue Proteins/biosynthesis,genetics Oocytes/ultrastructure Phosphoproteins/biosynthesis,genetics,immunology Phosphorylation Sequence Homology Tissue Distribution Xenopus Proteins Xenopus laevis/embryology,genetics
Chemicals
Actin Depolymerizing Factors Cofilin 1 Cofilin 2 Cytoskeletal Proteins DNA, Complementary Destrin Microfilament Proteins Nerve Tissue Proteins Phosphoproteins XAC1 protein, Xenopus XAC2 protein, Xenopus Xenopus Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Abe H
Department of Biology, Chiba University, Japan.
Obinata T
Minamide L S
Bamburg J R
References (52)
52 references, click to expand
  1. Calculation of protein extinction coefficients from amino acid sequence data.
    Anal Biochem. 1989 Nov 1;182(2):319-26 PMID: 2610349
  2. Nucleotide sequence and expression of a cDNA encoding chick brain actin depolymerizing factor.
    Biochemistry. 1990 Aug 14;29(32):7414-20 PMID: 2223773
  3. Sequence of cDNAs encoding actin depolymerizing factor and cofilin of embryonic chicken skeletal muscle: two functionally distinct actin-regulatory proteins exhibit high structural homology.
    Biochemistry. 1990 Aug 14;29(32):7420-5 PMID: 1699599
  4. A filter paper dye-binding assay for quantitative determination of protein without interference from reducing agents or detergents.
    Anal Biochem. 1990 Oct;190(1):66-70 PMID: 2285147
  5. Purification and characterization of low-molecular-weight actin-depolymerizing proteins from brain and cultured cells.
    Methods Enzymol. 1991;196:125-40 PMID: 1851937
  6. Actin-depolymerizing factor (ADF) in the cerebellum of the developing rat: a quantitative and immunocytochemical study.
    J Neurosci Res. 1991 Sep;30(1):18-27 PMID: 1795402
  7. Astrocyte process growth induction by actin breakdown.
    J Cell Biol. 1992 Apr;117(2):357-67 PMID: 1313815
  8. In situ hybridization: an improved whole-mount method for Xenopus embryos.
    Methods Cell Biol. 1991;36:685-95 PMID: 1811161
  9. Cofilin is an essential component of the yeast cortical cytoskeleton.
    J Cell Biol. 1993 Jan;120(2):421-35 PMID: 8421056
  10. Isolation of a yeast essential gene, COF1, that encodes a homologue of mammalian cofilin, a low-M(r) actin-binding and depolymerizing protein.
    Gene. 1993 Feb 14;124(1):115-20 PMID: 8440472
  11. On the crawling of animal cells.
    Science. 1993 May 21;260(5111):1086-94 PMID: 8493552
  12. Isolation and characterization of a regulated form of actin depolymerizing factor.
    J Cell Biol. 1993 Aug;122(3):623-33 PMID: 7687605
  13. Human actin depolymerizing factor mediates a pH-sensitive destruction of actin filaments.
    Biochemistry. 1993 Sep 28;32(38):9985-93 PMID: 8399167
  14. Analysis of the interactions of actin depolymerizing factor with G- and F-actin.
    Biochemistry. 1993 Sep 28;32(38):9994-10004 PMID: 8399168
  15. Occludin: a novel integral membrane protein localizing at tight junctions.
    J Cell Biol. 1993 Dec;123(6 Pt 2):1777-88 PMID: 8276896
  16. Green fluorescent protein as a marker for gene expression.
    Science. 1994 Feb 11;263(5148):802-5 PMID: 8303295
  17. The Caenorhabditis elegans unc-60 gene encodes proteins homologous to a family of actin-binding proteins.
    Mol Gen Genet. 1994 Feb;242(3):346-57 PMID: 8107682
  18. Ultrastructure of the yeast actin cytoskeleton and its association with the plasma membrane.
    J Cell Biol. 1994 Apr;125(2):381-91 PMID: 8163554
  19. Characterization and identification as cofilin and destrin of two thyrotropin- and phorbol ester-regulated phosphoproteins in thyroid cells.
    Exp Cell Res. 1994 May;212(1):49-61 PMID: 8174642
  20. Costimulatory signals for human T-cell activation induce nuclear translocation of pp19/cofilin.
    Proc Natl Acad Sci U S A. 1994 May 10;91(10):4494-8 PMID: 8183936
  21. Identification of Drosophila cytoskeletal proteins by induction of abnormal cell shape in fission yeast.
    Proc Natl Acad Sci U S A. 1994 May 10;91(10):4589-93 PMID: 8183953
  22. Characterization of a novel cofilin isoform that is predominantly expressed in mammalian skeletal muscle.
    J Biol Chem. 1994 May 27;269(21):15280-6 PMID: 8195165
  23. Dephosphorylation of cofilin in stimulated platelets: roles for a GTP-binding protein and Ca2+.
    Biochem J. 1994 Jul 1;301 ( Pt 1):41-7 PMID: 8037689
  24. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  25. An ultrastructural study of the first cleavage of Xenopus embryos.
    J Ultrastruct Res. 1974 Jun;47(3):433-51 PMID: 4365712
  26. High resolution two-dimensional electrophoresis of proteins.
    J Biol Chem. 1975 May 25;250(10):4007-21 PMID: 236308
  27. A general method for isolation of high molecular weight DNA from eukaryotes.
    Nucleic Acids Res. 1976 Sep;3(9):2303-8 PMID: 987581
  28. Albumin phylogeny for clawed frogs (Xenopus).
    Science. 1977 Feb 25;195(4280):785-7 PMID: 65013
  29. High resolution two-dimensional electrophoresis of basic as well as acidic proteins.
    Cell. 1977 Dec;12(4):1133-41 PMID: 23215
  30. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  31. Localization of thymosin beta 4 to the neural tissues during the development of Xenopus laevis, as studied by in situ hybridization and immunohistochemistry.
    Brain Res Dev Brain Res. 1994 Jun 17;79(2):177-85 PMID: 7955316
  32. Actin monomer binding proteins.
    Curr Opin Cell Biol. 1995 Feb;7(1):102-10 PMID: 7755981
  33. New horizons for cytokinesis.
    Curr Opin Cell Biol. 1995 Feb;7(1):23-31 PMID: 7755986
  34. The bulk of unpolymerized actin in Xenopus egg extracts is ATP-bound.
    Mol Biol Cell. 1995 Feb;6(2):227-36 PMID: 7787248
  35. Reactivation of phosphorylated actin depolymerizing factor and identification of the regulatory site.
    J Biol Chem. 1995 Jul 21;270(29):17582-7 PMID: 7615564
  36. Activation induces dephosphorylation of cofilin and its translocation to plasma membranes in neutrophil-like differentiated HL-60 cells.
    J Biol Chem. 1995 Aug 18;270(33):19551-6 PMID: 7642640
  37. Mutations in twinstar, a Drosophila gene encoding a cofilin/ADF homologue, result in defects in centrosome migration and cytokinesis.
    J Cell Biol. 1995 Dec;131(5):1243-59 PMID: 8522587
  38. The ADF/cofilin proteins: stimulus-responsive modulators of actin dynamics.
    Mol Biol Cell. 1995 Nov;6(11):1423-31 PMID: 8589446
  39. Partial purification and characterization of an actin depolymerizing factor from brain.
    FEBS Lett. 1980 Nov 17;121(1):178-82 PMID: 6893966
  40. A simple method of reducing the fading of immunofluorescence during microscopy.
    J Immunol Methods. 1981;43(3):349-50 PMID: 7019347
  41. The depolymerization of actin by specific proteins from plasma and brain: a quantitative assay.
    Anal Biochem. 1982 Jan 1;119(1):102-14 PMID: 7072932
  42. A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids.
    Anal Biochem. 1984 Apr;138(1):141-3 PMID: 6731838
  43. Cofilin, a protein in porcine brain that binds to actin filaments and inhibits their interactions with myosin and tropomyosin.
    Biochemistry. 1984 Oct 23;23(22):5307-13 PMID: 6509022
  44. pH control of actin polymerization by cofilin.
    J Biol Chem. 1985 Nov 25;260(27):14410-2 PMID: 4055781
  45. Kinematics of gray crescent formation in Xenopus eggs: the displacement of subcortical cytoplasm relative to the egg surface.
    Dev Biol. 1986 Feb;113(2):484-500 PMID: 3949075
  46. Reversible induction of actin rods in mouse C3H-2K cells by incubation in salt buffers and by treatment with non-ionic detergents.
    Exp Cell Res. 1986 Jun;164(2):492-506 PMID: 3086110
  47. Actin and actin-binding proteins. A critical evaluation of mechanisms and functions.
    Annu Rev Biochem. 1986;55:987-1035 PMID: 3527055
  48. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
    Anal Biochem. 1987 Apr;162(1):156-9 PMID: 2440339
  49. Subcortical rotation in Xenopus eggs: an early step in embryonic axis specification.
    Dev Biol. 1987 Oct;123(2):526-39 PMID: 3653523
  50. Distribution among tissues and intracellular localization of cofilin, a 21kDa actin-binding protein.
    Cell Struct Funct. 1987 Oct;12(5):443-52 PMID: 3315240
  51. Distribution and cellular localization of actin depolymerizing factor.
    J Cell Biol. 1987 Dec;105(6 Pt 1):2817-25 PMID: 3320057
  52. Dephosphorylation of cofilin accompanies heat shock-induced nuclear accumulation of cofilin.
    J Biol Chem. 1989 Sep 25;264(27):16143-8 PMID: 2777782
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.132.5.871
Published
1996-03-00
Pages
871-85
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120733
Subset
IM
Grants
NIGMS NIH HHS · GM35126 · United States
Databases
GENBANK
U26269, U26270
External Links
PubMed source DOI: 10.1083/jcb.132.5.871 PMC Full Text View journal details
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