Towards a molecular understanding of cytokinesis

doi:10.1016/S0962-8924(00)01747-5

Trends in Cell Biology

Volume 10, Issue 6, 1 June 2000, Pages 228-237

https://doi.org/10.1016/S0962-8924(00)01747-5 Get rights and content

Abstract

In this review, we focus on recent discoveries regarding the molecular basis of cleavage furrow positioning and contractile ring assembly and contraction during cytokinesis. However, some of these mechanisms might have different degrees of importance in different organisms. This synthesis attempts to uncover common themes and to reveal potential relationships that might contribute to the biochemical and mechanical aspects of cytokinesis. Because the information about cytokinesis is still fairly rudimentary, our goal is not to present a definitive model but to present testable hypotheses that might lead to a better mechanistic understanding of the process.

Section snippets

Spatio-temporal control by microtubules

Since the classic experiments of the early twentieth century (reviewed in Ref. 2), the astral microtubules have been implicated in determining the site of cleavage. In a more recent experiment, Rappaport² manipulated echinoderm eggs using a glass bead to generate binucleate, horseshoe-shaped cells with a spindle in each arm of the cell. These cells divided between asters of the same spindle and between the asters of adjacent spindles. In recent years, these findings have been extended to other

Other spatio-temporal controllers

Mutagenesis experiments in S. pombe have produced several cytokinesis mutants²⁶. Three of these, spg1, mid1 and cdc15, have phenotypes that indicate they are very important for the spatio-temporal control of the formation of the contractile ring. The Spg1p is a Ras superfamily small GTPase²⁷ that is localized to the spindle pole body²⁸. Loss of this protein leads to a failure in septum formation, whereas overexpression of it can induce septum formation during other times of the cell cycle²⁷.

Cortical tension and flow

Most models for animal cytokinesis include a role for the generation of a cortical-tension differential with the peak of tension at the cell equator and the nadir of tension at the cell poles (reviewed in Ref. 2). This cortical-tension differential is thought to contribute to two aspects of cytokinesis: the assembly of the contractile ring by generating cortical flow of contractile ring elements towards the furrow region; and the constriction of the cleavage furrow generated by the actomyosin

Rho-mediated signalling events

During cytokinesis, Rho-mediated signalling is required for the regulation of cortical activities during cytokinesis46, 47 in addition to the recruitment of myosin II and actin filaments to the cleavage furrow⁴⁸. The D. melanogaster pebble that encodes an ECT2-family guanine nucleotide exchange factor that is specific for Rho1 is also required for cytokinesis and is localized to the contractile ring similar to anillin (discussed below) and the Drosophila peanut (a septin)⁴⁷. pebble mutants fail

Anchoring the contractile ring to the membrane

Although it is not known exactly how the contractile ring is anchored to the plasma membrane, some proteins that are involved in anchoring the cytoskeleton to the plasma membrane are localized to the cleavage furrow. The CD43 integral membrane protein⁵⁴ and radixin⁵⁵ localize to the cleavage furrow in mammalian cells. The cytoplasmic domain of CD43 can interact with the N-terminal, globular domain of radixin, that in turn binds to filamentous actin, thereby crosslinking the actin to the plasma

Assembly of actin-associated proteins

In most systems, a contractile ring contains a band of actin filaments. This band of actin filaments is well organized into antiparallel arrays of microfilaments. Alternatively, in cells such as D. discoideum, it is more diffusely distributed along the equatorial zone. Both pre-existing filaments and new actin-filament polymerization contribute to the formation of the contractile ring actin61, 62. However, genetic studies from a variety of organisms suggest that either of these two mechanisms

Cleavage furrow contraction

Nonmuscle myosin II has been classically thought of as the motor protein that drives purse-string cleavage of the cell. It is localized to the cleavage furrows from S. pombe to mammals and it is thought to provide the contractile force of the cleavage furrow. Genetic evidence from several organisms, including D. discoideum (see for example 41, 83), C. elegans⁸⁴ and S. pombe85, 86, has suggested a general requirement for myosin II in cytokinesis.

However, in D. discoideum, all of the mutant cell

Regulation of the myosin II-mediated contractility

By using a biosensor for the myosin II regulatory light-chain activating phosphorylation on serine-19, DeBiasio et al.³⁷ demonstrated that there is a global rise in phosphorylation of myosin II regulatory light chain by anaphase of mammalian cells. Later, this phosphorylation becomes somewhat spatially restricted to the region of the cleavage furrow by telophase, but this phosphorylation decreases through cytokinesis. Indeed, a requirement for regulatory light chain kinase has been indicated in

Membrane dynamics

To increase the surface area of the cell as it divides, additional membrane might be recruited into the plasma membrane. At least some of this recruitment comes from the insertion of vesicles at the sight of the invaginating cleavage furrow⁴⁸. Syntaxins are t-SNAREs that are required on the target membrane to promote fusion of the vesicle with the target membrane during exocytosis. Recently, mutations in two distantly related syntaxin-encoding genes, the Arabidopsis thaliana Knolle1¹⁰⁸ and the

Completion of cytokinesis

Several new proteins and signalling events are required for the final severing of the midbody (Fig. 1, 5:00). Loss-of-function mutant D. discoideum strains in genes that encode a Ras isoform¹¹⁵, an IQGAP isoform¹¹⁶ and calmodulin¹¹⁷ fail at this late step of cytokinesis. The cells are able to propagate themselves on surfaces because the daughter cells can crawl away from each other until the unsevered midbody finally breaks. In C. elegans, the formin protein, cyk-1, appears to be required for a

Conclusion

Cytokinesis involves nearly every field of cell biology, including signal transduction, membrane trafficking, integral-membrane and peripheral-cytoskeletal proteins, myosin mechanochemistry and general cellular physiology. Many of the molecular components are being revealed through genetic screens in model systems such as D. discoideum, S. pombe, S. cerevisiae, C. elegans and D. melanogaster. Although certain themes might not be entirely universal, some mechanisms are emerging as being used

Acknowledgements

We thank Ji-Hong Zang for the GFP–myosin II movie series used to generate Fig. 1. We thank all the members of the Spudich laboratory for numerous useful discussions. This work was supported by a Runyon–Winchell fellowship #DRG1457 to D.N.R. and an NIH grant #GM40509 to J.A.S.

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Trends in Cell Biology

ReviewTowards a molecular understanding of cytokinesis

Abstract

Section snippets

Spatio-temporal control by microtubules

Other spatio-temporal controllers

Cortical tension and flow

Rho-mediated signalling events

Anchoring the contractile ring to the membrane

Assembly of actin-associated proteins

Cleavage furrow contraction

Regulation of the myosin II-mediated contractility

Membrane dynamics

Completion of cytokinesis

Conclusion

Acknowledgements

Trends Cell Biol.

Curr. Opin. Cell Biol.

Trends Cell Biol.

Curr. Biol.

Cell

Cell

J. Biol. Chem.

Biophys. J.

Cell

Cell

Cell

Exp. Cell Res.

Cell

J. Biol. Chem.

Curr. Opin. Cell Biol.

Cytokinesis in Animal Cells

Unusual cleavage furrows in vertebrate tissue culture cells: insights into the mechanisms of cytokinesis

Cell Motil. Cytoskeleton

Microtubule-mediated centrosome motility and the positioning of cleavage furrows in multinucleate myosin II-null cells

J. Cell Sci.

Interpolar spindle microtubules in PTK cells

J. Cell Biol.

Signals from the spindle midzone are required for the stimulation of cytokinesis in cultured epithelial cells

Mol. Biol. Cell

Midzone microtubule bundles are continuously required for cytokinesis in cultured epithelial cells

J. Cell Biol.

Cooperative interactions between the central spindle and the contractile ring during Drosophila cytokinesis

Genes Dev.

A plus-end-directed motor enzyme that moves antiparallel microtubules in vitro localizes to the interzone of mitotic spindles

Nature

Cytokinesis and midzone microtubule organization in Caenorhabditis elegans require the kinesin-like protein zen-4

Mol. Biol. Cell

pavarotti encodes a kinesin-like protein required to organize the central spindle and contractile ring for cytokinesis

Genes Dev.

Cleavage furrows formed between centrosomes lacking an intervening spindle and chromosomes contain microtubule bundles, INCENP, and CHO1 but not CENP-E

Mol. Biol. Cell

The conserved Schizosaccharomyces pombe kinase plo1, required to form a bipolar spindle, the actin ring, and septum, can drive septum formation in G1 and G2 cells

Genes Dev.

Drosophila polo kinase is required for cytokinesis

J. Cell Biol.

Plk is an M-phase specific protein kinase and interacts with a kinesin-like protein, CHO1/MKLP-1

Mol. Cell. Biol.

Displacement of cleavage plane in the sea urchin egg by locally applied taxol

Cell Motil. Cytoskeleton

Inhibition of chromosomal separation provides insights into cleavage furrow stimulation in cultured epithelial cells

Mol. Biol. Cell

Targeting, capture, and stabilization of microtubules at early focal adhesions

J. Cell Biol.

Talin-null cells of Dictyostelium are strongly defective in adhesion to particle and substrate surfaces and slightly impaired in cytokinesis

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Occurrence of fibers and their association with talin in the cleavage furrows of PtK2 cells

Cell Motil. Cytoskeleton

Microtubule growth activates Rac1 to promote lamellipodial protrusion in fibroblasts

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Parameters that specify the timing of cytokinesis

J. Cell Biol.

Isolation and characterization of fission yeast mutants defective in the assembly and placement of the contractile actin ring

J. Cell Sci.

The Spg1p GTPase is an essential, dosage-dependent inducer of septum formation in Schizosaccharomyces pombe

Genes Dev.

Asymmetric segregation on spindle poles of the Schizosaccharomyces pombe septum-inducing protein kinase Cdc7p

Genes Dev.

Role of polo kinase and mid1p in determining the site of cell division in fission yeast

J. Cell Biol.

Review
Towards a molecular understanding of cytokinesis

The conserved Schizosaccharomyces pombe kinase plo1, required to form a bipolar spindle, the actin ring, and septum, can drive septum formation in G₁ and G₂ cells