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A possible role for actin dots in the formation of the contractile ring in the ultra-micro algaCyanidium caldarium RK-1

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Summary

In the primitive red algaCyanidium caldarium RK-1, cytokinesis is controlled by a simple contractile ring, as in animal cells. To clarify the mechanism of formation of the contractile ring, we isolated actin genes and performed an immunocytological study.C. caldarium RK-1 has two actin genes encoding proteins with the same sequence of 377 amino acids. The primary structure indicated that the actin molecules ofC. caldarium RK-1 are typical, despite the fact that the organism is considered to be phylogenetically primitive. We prepared antiserum against aC. caldarium RK-1 actin fusion protein and indirect immunofluorescence staining was performed. In interphase cells, many actin dots were observed in the cytoplasm but none at the future cleavage plane. Prior to cytokinesis, some of these dots appeared and became aligned along the equatorial plane. At the same time, a thin “immature” contractile ring was observed to appear to be formed by connection of the aligned actin dots. This immature contractile ring thickened to nearly its maximum size by the time cytokinesis began. The formation of the contractile ring seemed to be a result of de novo assembly of actin monomers, rather than a result of the accumulation and bundling of pre-existing actin filaments. During the constriction of the contractile ring, no actin dots were observed in the cytoplasm. These observations suggest that actin dots are responsible for the formation of the contractile ring, but are not necessary for its disintegration. Furthermore, immunogold localization specific for actin revealed at electron microscopy level that fine filaments running just beneath the cleavage furrow are, in fact, actin filaments.

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Abbreviations

ORF:

open reading frame

IPTG:

isopropyl-β-D(−)-thiogalactopyranoside

SDS-PAGE:

sodium dodecyl sulphate-poly-acrylamide gel electrophoresis

DAPI:

4′,6-diamidino-2-phenylindole

References

  • Adams AEM, Pringle JR (1984) Relationship of actin and tubulin distribution to bud growth in wild-type and morphogeneticmutantSaccharomyces cerevisiae. J Cell Biol 98: 934–945

    PubMed  Google Scholar 

  • Allen MB (1959) Studies withCyanidium caldarium, an anomalously pigmented chlorophyte. Arch Microbiol 32: 270–277

    Google Scholar 

  • Baskin TI, Cande WZ (1991) The structure and function of the mitotic spindle in flowering plants. Annu Rev Plant Physiol Plant Mol Biol 41: 277–315

    Google Scholar 

  • Blake CCF (1978) Exons encode protein functional units. Nature 277: 598

    Google Scholar 

  • — (1983) Exons: present from the beginning? Nature 306: 535–537

    PubMed  Google Scholar 

  • Cande WZ (1980) A permeabilized cell model for studying cytokinesis using mammalian tissue culture cells. J Cell Biol 87: 326–335

    PubMed  Google Scholar 

  • Cao LG, Wang YL (1990a) 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 110: 1089–1096

    PubMed  Google Scholar 

  • — — (1990b) Mechanism of the formation of contractile ring in dividing cultured animal cells II: cortical movement of microinjected actin filaments. J Cell Biol 111: 1905–1912

    PubMed  Google Scholar 

  • Cavalier-Smith T (1985) Selfish DNA and the origin of introns. Nature 315: 283–284

    PubMed  Google Scholar 

  • —, (1991) Intron phylogeny: a new hypothesis. Trends Genet 7: 145–148

    PubMed  Google Scholar 

  • Clayton L, Lloyd CW (1985) Actin organization during the cell cycle in meristematic plant cells. Exp Cell Res 156: 231–238

    PubMed  Google Scholar 

  • Cresnar B, Mages W, Müller K, Salbaum JM, Schmitt R (1990) Structure and expression of a single actin gene inVolvox carteri. Curr Genet 18: 337–346

    PubMed  Google Scholar 

  • Darnell JE (1978) Implications of RNA · RNA splicing in evolution of eukaryotic cells. Science 202: 1257–1260

    PubMed  Google Scholar 

  • De Lozanne A, Spudich JA (1987) Disruption of theDictyostelium myosin heavy chain gene by homologous recombination. Science 236: 1086–1091

    PubMed  Google Scholar 

  • Doolittle WF (1978) Genes in pieces: were they ever together? Nature 272: 581–582

    Google Scholar 

  • Drubin DG, Jones HD, Wertman KF (1993) Actin structure and function: roles in mitochondrial organization and morphogenesis in budding yeast and identification of the phalloidin-binding site. Mol Biol Cell 4: 1277–1294

    PubMed  Google Scholar 

  • Erba HP, Eddy R, Shows T, Kedes L, Gunning P (1988) Structure, chromosome location, and expression of the human gammaactin gene: differential evolution, location, and expression of the cytoskeletal beta- and gamma-actin genes. Mol Cell Biol 8: 1775–1789

    PubMed  Google Scholar 

  • Fidel S, Doonan JH, Morris NR (1988)Aspergillus nidulans contains a single actin gene which has unique intron locations and encodes a γ-actin. Gene 70: 283–293

    PubMed  Google Scholar 

  • Fishkind DJ, Wang YL (1995) New horizons for cytokinesis. Curr Opin Cell Biol 7: 23–31

    PubMed  Google Scholar 

  • Fornwald JA, Kuncio G, Peng I, Ordahl CP (1982) The complete nucleotide sequence of the chick α-actin gene and its evolutionary relationship to the actin gene family. Nucleic Acids Res 10: 3861–3876

    PubMed  Google Scholar 

  • Fujie M, Kuroiwa H, Kawano S, Mutoh S, Kuroiwa T (1994) Behavior of organelles and their nucleoids in the shoot apical meristem during leaf development inArabidopsis thaliana L. Planta 194: 395–405

    Google Scholar 

  • Fyrberg EA, Mahaffey JW, Bond BJ, Davidson N (1983) Transcripts of the sixDrosophila actin genes accumulate in a stage- and tissue-specific manner. Cell 33: 115–123

    PubMed  Google Scholar 

  • Gallwitz D, Sures I (1980) Structure of a split yeast gene: complete nucleotide sequence of the actin gene inSaccharomyces cerevisiae. Proc Natl Acad Sci USA 77: 2546–2550

    PubMed  Google Scholar 

  • Hamada H, Petrino MG, Kakunaga T (1982) Molecular structure and evolutionary origin of human cardiac muscle actin gene. Proc Natl Acad Sci USA 79: 5901–5905

    PubMed  Google Scholar 

  • Harper DS, Jahn CL (1989) Differential use of termination codons in ciliated protozoa. Proc Natl Acad Sci USA 86: 3252–3256

    PubMed  Google Scholar 

  • Hirono M, Endoh H, Okada N, Numata O, Watanabe Y (1987)Tetrahymena actin: cloning and sequencing of theTetrahymena actin gene and identification of its gene product. J Mol Biol 194: 181–192

    PubMed  Google Scholar 

  • —, Kumagai Y, Numata O, Watanabe Y (1989) Purification ofTetrahymena actin reveals some unusual properties. Proc Natl Acad Sci USA 86: 75–79

    PubMed  Google Scholar 

  • Katoh T, Morita F (1995) Mapping myosin-binding sites on actin probed by peptides that inhibit actomyosin interaction. J Biochem 120: 580–586

    Google Scholar 

  • Kilmartin JV, Adams AEM (1984) Structural rearrangements of tubulin and actin during the cell cycle of the yeastSaccharomyces. J Cell Biol 98: 922–933

    Google Scholar 

  • Knecht DA, Loomis WF (1987) Antisense RNA inactivation of myosin heavy chain gene expression inDictyostelium discoideum. Science 236: 1086–1091

    PubMed  Google Scholar 

  • Kusakabe T, Hikosaka A, Satoh N (1995) Coexpression and promoter function in two muscle actin gene complexes of different structural organization in the ascidianHalocynthia roretzi. Dev Biol 169: 461–472

    PubMed  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227: 680–685

    PubMed  Google Scholar 

  • Mabuchi I (1986) Biochemical aspects of cytokinesis. Int Rev Cytol 101: 175–213

    PubMed  Google Scholar 

  • — (1994) Cleavage furrow: timing of emergence of contractile ring actin filaments and establishment of the contractile ring by filament bundling in sea urchin eggs. J Cell Sci 107: 1853–1862

    PubMed  Google Scholar 

  • —, Okuno M (1977) The effect of myosin antibody on the division of the starfish blastomeres. J Cell Biol 74: 251–263

    Google Scholar 

  • —, Tsukita Sh, Tsukita Sa, Sawai T (1988) Cleavage furrow isolated from newt eggs: contraction, organization of the actin filaments, and protein components of the furrow. Proc Natl Acad Sci USA 85: 5966–5970

    PubMed  Google Scholar 

  • Melloul D, Aloni B, Calvo J, Yaffe D, Nudel U (1984) Developmentally regulated expression of chimeric genes containing muscle actin DNA sequences in transfected myogenic cells. EMBO J 3: 983–990

    PubMed  Google Scholar 

  • Mertins P, Gallwitz D (1987) A single intronless actin gene in the fission yeastSchizosaccharomyces pombe: nucleotide sequence and transcripts formed in homologous and heterologous yeast. Nucleic Acids Res 15: 7369–7379

    PubMed  Google Scholar 

  • Miki M, Barden JA, dos Remedios CG, Phillips L, Hambly BD (1987) Interaction of phalloidin with chemically modified actin. Eur J Biochem 165: 125–130

    PubMed  Google Scholar 

  • Mita T, Kuroiwa T (1988) Division of plastids by a plastid-dividing ring inCyanidium caldarium. Protoplasma Suppl 1: 133–152

    Google Scholar 

  • —, Kanbe T, Tanaka T, Kuroiwa T (1986) A ring structure around the dividing plane of theCyanidium caldarium chloroplast. Protoplasma 130: 211–213

    Google Scholar 

  • Mounier N, Prudhomme JC (1991) Differential expression of muscle and cytoplasmic actin genes during development ofBombyx mori. Insect Biochem 21: 523–533

    Google Scholar 

  • Nagashima H, Fukuda I (1981) Morphological properties ofCyanidium caldarium and related algae in Japan. Jpn J Phycol 29: 237–242

    Google Scholar 

  • Nairn CJ, Winesett L, Ferl RJ (1988) Nucleotide sequence of an actin gene fromArabidopsis thaliana. Gene 65: 247–257

    PubMed  Google Scholar 

  • Ng R, Abelson JN (1980) Isolation and sequence of the gene for actin inSaccharomyces cerevisiae. Proc Natl Acad Sci USA 77: 3912–3916

    PubMed  Google Scholar 

  • Ohta N, Nagashima H, Kawano S, Kuroiwa T (1992) Isolation of the chloroplast DNA and the sequence of thetrnK gene fromCyanidium caldarium strain RK-1. Plant Cell Physiol 33: 657–661

    Google Scholar 

  • —, Sato N, Kawano S, Kuroiwa T (1994) ThetrpA gene on the plastid genome ofCyanidium caldarium strain RK-1. Curr Genet 25: 357–361

    PubMed  Google Scholar 

  • Rappaport R (1986) Establishment of the mechanism of cytokinesis in animal cells. Int Rev Cytol 105: 245–281

    PubMed  Google Scholar 

  • Remedies CGD, Moens PDJ (1995) Actin and the actomyosin interface: a review. Biochim Biophys Acta 1228: 99–124

    PubMed  Google Scholar 

  • Romans P, Firtel RA (1985) Organization of the actin multigene family ofDictyostelium discoideum and analysis of variability in the protein coding regions. J Mol Biol 186: 321–335

    PubMed  Google Scholar 

  • Sanchez F, Tobin SL, Rdest U, Zulauf E, McCarthy BJ (1983) TwoDrosophila actin genes in detail: gene structure, protein structure and transcription during development. J Mol Biol 163: 533–551

    PubMed  Google Scholar 

  • Sanger JM, Sanger JW (1980) Banding and polarity of actin filaments in interphase and cleaving cells. J Cell Biol 86: 568–575

    Google Scholar 

  • Satterwhite LL, Pollard TD (1992) Cytokinesis. Curr Opin Cell Biol 4: 43–52

    PubMed  Google Scholar 

  • Schmit AC, Lambert AM (1990) Microinjected fluorescent phalloidin in vivo reveals the F-actin dynamics and assembly in higher plant mitotic cells. Plant Cell 2: 129–138

    PubMed  Google Scholar 

  • Schroeder TE (1968) Cytokinesis: filaments in the cleavage furrow. Exp Cell Res 53: 272–276

    PubMed  Google Scholar 

  • — (1970) The contractile ring I: fine structure of dividing mammalian (HeLa) cells and the effects of cytochalasin B. Z Zellforsch Mikrosk Anat 109: 431–449

    PubMed  Google Scholar 

  • — (1972) The contractile ring II: determining its brief existence, volumetric changes, and vital role in cleavingArbacia eggs. J Cell Biol 53: 419–434

    PubMed  Google Scholar 

  • — (1975) Dynamics of the contractile ring. In: Inoué S, Stephens RE (eds) Molecules and cell movement. Raven, New York, pp 305–334

    Google Scholar 

  • Seckbach J (1996) Biological aspects of the origin of life: open questions in eukaryogenesis. In: Flores JC, Raulin F (eds) Chemical evolution: physics of the origin and evolution of life. Kluwer, Dordrecht, pp 197–213

    Google Scholar 

  • —, González E, Wainwright IM, Gross W (1992) Peroxisomal function in the Cyanidiophyceae (Rhodophyta): a discussion of phylogenetic relationships and the evolution of microbodies (peroxisomes). Nova Hedwigia 55: 99–109

    Google Scholar 

  • Selman GG, Perry MM (1970) Ultrastructural changes in the surface layers of the newt's egg in relation to the mechanism of its cleavage. J Cell Sci 6: 207–227

    PubMed  Google Scholar 

  • Sugase Y, Hirono M, Kindle KL, Kamiya, R (1996) Cloning and characterization of the actin-encoding gene ofChlamydomonas reinhardtii. Gene 168: 117–121

    PubMed  Google Scholar 

  • Suzuki K, Ohta N, Kuroiwa T (1992) Isolation of the cell-nuclear, mitochondrial, and chloroplast DNA from the ultra-small eukaryoteCyanidioschyzon merolae. Protoplasma 171: 80–84

    Google Scholar 

  • —, Kawazu T, Mit T, Takahashi H, Itoh R, Toda K, Kuroiwa T (1995) Cytokinesis by a contractile ring in the primitive red algaCyanidium caldarium RK-1. Eur J Cell Biol 67: 170–178

    PubMed  Google Scholar 

  • Szollosi D (1970) Cortical cytoplasmic filaments of cleaving eggs: a structural element corresponding to the contractile ring. J Cell Biol 44: 192–209

    PubMed  Google Scholar 

  • Takahashi H, Takano H, Yokoyama A, Hara Y, Kawano S, Toh-e A, Kuroiwa T (1995) Isolation, characterization and chromosomal mapping of an actin gene from the primitive red algaCyanidioschyzon merolae. Curr Genet 28: 484–490

    PubMed  Google Scholar 

  • —, Kuroiwa H, Miyagishima S, Toda K, Itoh R, Kuroiwa T (1997) Improved procedure for immunogold electron microscopy: rapid-freeze substitution with absolute acetone. Cytologia 62: 303–308

    Google Scholar 

  • Tilney LG, Marsland D (1969) A fine structural analysis of cleavage induction and furrowing in the eggs ofArbacia punctulata. J Cell Biol 42: 170–184

    PubMed  Google Scholar 

  • Usui N, Yoneda M (1982) Ultrastructural basis of the tension increase in sea urchin eggs prior to cytokinesis. Dev Growth Differ 24: 453–465

    Google Scholar 

  • Vandekerckhove J, Deboben A, Nassal M, Wieland T (1985) The phalloidin binding site of F-actin. EMBO J 4: 2815–2818

    PubMed  Google Scholar 

  • Weber K, Kabsch W (1994) Intron positions in actin genes seem unrelated to the secondary structure of the protein. EMBO J 13: 1280–1286

    PubMed  Google Scholar 

  • Wick SM (1991) Spatial aspect of cytokinesis in plant cells. Curr Opin Cell Biol 3: 253–260

    PubMed  Google Scholar 

  • Wieland T (1977) Modification of actins by phallotoxins. Naturwissenschaften 64: 303–309

    PubMed  Google Scholar 

  • Wulf E, Deboden A, Bautz FA, Faulstich H, Wieland TH (1979) Fluorescent phallotoxin, a tool for the visualization of cellular actin. Proc Natl Acad Sci USA 76: 4498–4502

    PubMed  Google Scholar 

  • Yumura S (1996) Spatial distribution of fluorescently labeled actin in livingDictyostelium amoebae. Cell Struct Funct 21: 189–197

    PubMed  Google Scholar 

  • Zang D, Wadsworth P, Hepler PK (1993) Dynamics of microfilaments are similar, but distinct from microtubules during cytokinesis in living, dividing plant cells. Cell Motil Cytoskeleton 24: 151–155

    Google Scholar 

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Authors and Affiliations

  1. Department of Biological Sciences, Graduate School of Science, University of Tokyo, Hongo, 113-0033, Tokyo, Japan

    H. Takahashi, H. Takano, R. Itoh, K. Toda, S. Kawano & T. Kuroiwa

  2. Kyoritsu Women's Junior College, Kanda, Tokyo

    H. Kuroiwa

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  1. H. Takahashi
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  2. H. Takano
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  3. H. Kuroiwa
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Takahashi, H., Takano, H., Kuroiwa, H. et al. A possible role for actin dots in the formation of the contractile ring in the ultra-micro algaCyanidium caldarium RK-1. Protoplasma 202, 91–104 (1998). https://doi.org/10.1007/BF01280878

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