To study centrosome motility and the interaction of microtubules with the cell cortex in mitotic, post-mitotic and interphase cells, (alpha)-tubulin was tagged in Dictyostelium discoideum with green fluorescent protein. Multinucleate cells formed by myosin II-null mutants proved to be especially suited for the analysis of the control of cleavage furrow formation by the microtubule system. After docking of the mitotic apparatus onto the cell cortex during anaphase, the cell surface is activated to form ruffles on top of the asters of microtubules that emanate from the centrosomes. Cleavage furrows are initiated at spaces between the asters independently of the positions of spindles. Once initiated, the furrows expand as deep folds without a continued connection to the microtubule system. Occurrence of unilateral furrows indicates that a closed contractile ring is dispensable for cytokinesis in Dictyostelium. The progression of cytokinesis in the multinucleate cells underlines the importance of proteins other than myosin II in specifying a cleavage furrow. The analysis of centrosome motility suggests a major role for a minus-end directed motor protein, probably cytoplasmic dynein, in applying traction forces on guiding microtubules that connect the centrosome with the cell cortex.

Adachi
H.
,
Takahashi
Y.
,
Hasebe
T.
,
Shirouzu
M.
,
Yokoyama
S.
,
Sutoh
K.
(
1997
).
Dictyostelium IQGAP-related protein specifically involved in the completion of cytokinesis.
J. Cell Biol
137
,
891
898
Amos
L. A.
,
Amos
W. B.
(
1991
).
The bending of sliding microtubules imaged by confocal light microscopy and negative stain electron microscopy.
J. Cell Sci
14
,
95
101
Best
A.
,
Ahmed
S.
,
Kozma
R.
,
Lim
L.
(
1996
).
The Ras-related GTPase Rac1 binds tubulin.
J. Biol. Chem
271
,
3756
3762
Burton
K.
,
Taylor
D. L.
(
1997
).
Traction forces of cytokinesis measured with optically modified elastic substrata.
Nature
385
,
450
454
Cao
L.-G.
,
Wang
Y.-L.
(
1996
).
Signals from the spindle midzone are required for the stimulation of cytokinesis in cultured epithelial cells.
Mol. Biol. Cell
7
,
225
232
Carminati
J. L.
,
Stearns
T.
(
1997
).
Microtubules orient the mitotic spindle in yeast through dynein-dependent interactions with the cell cortex.
J. Cell Biol
138
,
629
641
Chenn
A.
,
McConnell
S. K.
(
1995
).
Cleavage orientation and the asymmetric inheritance of Notch 1 immunoreactivity in mammalian neurogenesis.
Cell
82
,
631
641
Conrad
G. W.
,
Schroeder
T. E.
(
1990
).
Cytokinesis: mechanisms of furrow formation during cell division.
Ann. NY Acad. Sci
582
,
327
–.
Cramer
L. P.
,
Mitchison
T. J.
(
1995
).
Myosin is involved in postmitotic cell spreading.
J. Cell Biol
131
,
179
189
DeLozanne
A.
,
Spudich
J. A.
(
1987
).
Disruption of the Dictyostelium myosin heavy chain gene by homologous recombination.
Science
236
,
1086
1091
Devore
J. J.
,
Conrad
G. W.
,
Rappaport
R.
(
1989
).
A model for astral stimulation of cytokinesis in animal cells.
J. Cell Biol
109
,
2225
2232
Euteneuer
U.
,
Schliwa
M.
(
1985
).
Evidence for an involvement of actinin the positioning and motility of centrosomes.
J. Cell Biol
101
,
96
103
Faix
J.
,
Dittrich
W.
(
1996
).
DGAP1, a homologue of rasGTPase activating proteins that controls growth, cytokinesis and development in Dictyostelium discoideum.
FEBS Lett
394
,
251
257
Faix
J.
,
Steinmetz
M.
,
Boves
H.
,
Kammerer
R. A.
,
Lottspeich
F.
,
Mintert
U.
,
Murphy
J.
,
Stock
A.
,
Aebi
U.
,
Gerisch
G.
(
1996
).
Cortexillins, major determinants of cell shape and size, are actin-bundling proteins with a parallel coiled-coil tail.
Cell
86
,
631
642
Felgner
H.
,
Frank
R.
,
Schliwa
M.
(
1996
).
Flexural rigidity of microtubules measured with the use of optical tweezers.
J. Cell Sci
109
,
509
516
Gittes
F.
,
Mickey
B.
,
Nettleton
J.
,
Howard
J.
(
1993
).
Flexural rigidity of microtubules and actin filaments measured from thermal fluctuations in shape.
J. Cell Biol
120
,
923
934
Hara
K.
,
Tydeman
P.
,
Kirschner
M.
(
1980
).
A cytoplasmic clock with the same period as the division cycle in Xenopus eggs.
Proc. Nat. Acad. Sci USA
77
,
462
466
Harris
A. K.
,
Gewalt
S. L.
(
1989
).
Simulation testing of mechanisms for inducing the formation of the contractile ring in cytokinesis.
J. Cell Biol
109
,
2215
2223
He
X.
,
Dembo
M.
(
1997
).
On the mechanics of the first cleavage division of the sea urchin egg.
Exp. Cell Res
233
,
252
273
Heim
R.
,
Tsien
R. Y.
(
1996
).
Engineering green fluorescent protein for improved brightness, longer wavelength and fluorescence resonance energy transfer.
Curr. Biol
6
,
178
182
Hyman
A. A.
(
1989
).
Centrosome movement in the early divisions of Caenorhabditis elegans: A cortical site determining centrosome position.
J. Cell Biol
109
,
1185
1193
Hyman
A. A.
,
White
J. G.
(
1987
).
Determination of cell division axes in the early embryogenesis of Caenorhabditis elegans.
J. Cell Biol
105
,
2123
2135
Jung
G.
,
Wu
X.
,
Hammer
J. A.
III
(
1996
).
Dictyostelium mutants lacking multiple classic myosin I isoforms reveal combinations of shared and distinct functions.
J. Cell Biol
133
,
305
323
Koonce
M. P.
,
Grissom
P. M.
,
McIntosh
J. R.
(
1992
).
Dynein from Dictyostelium: primary structure comparisons between a cytoplasmic motor enzyme and flagellar dynein.
J. Cell Biol
119
,
1597
1604
Koonce
M. P.
,
McIntosh
J. R.
(
1990
).
Identification and immunolocalization of cytoplasmic dynein in Dictyostelium.
Cell Mot. Cytoskel
15
,
51
62
Koonce
M. P.
,
Samso
M.
(
1996
).
Overexpression of cytoplasmic dynein's globular head causes a collapse of the interphase microtubule network in Dictyostelium.
Mol. Biol. Cell
7
,
935
948
Larochelle
D. A.
,
Vithalani
K. K.
,
DeLozanne
A.
(
1996
).
A novel member of the rho family of small GTP-binding proteins is specifically required for cytokinesis.
J. Cell Biol
133
,
1321
1329
Manstein
D. J.
,
Titus
M. A.
,
DeLozanne
A.
,
Spudich
J. A.
(
1989
).
Gene replacement in Dictyostelium: generation of myosin null mutants.
EMBO J
8
,
923
932
McIntosh
J. R.
,
Roos
U.-P.
,
Neighbors
B.
,
McDonald
K. L.
(
1985
).
Architecture of the microtubule component of mitotic spindles from Dictyostelium discoideum.
J. Cell Sci
75
,
93
129
Mickey
B.
,
Howard
J.
(
1995
).
Rigidity of microtubules is increased by stabilizing agents.
J. Cell Biol
130
,
909
917
Moens
P. B.
(
1976
).
Spindle and kinetochore morphology of Dictyostelium discoideum.
J. Cell Biol
68
,
113
122
Neujahr
R.
,
Heizer
C.
,
Gerisch
G.
(
1997
).
Myosin II-independent processes in mitotic cells of Dictyostelium discoideum: redistribution of the nuclei, re-arrangement of the actin system and formation of the cleavage furrow.
J. Cell Sci
110
,
123
137
Neujahr
R.
,
Heizer
C.
,
Albrecht
R.
,
Ecke
M.
,
Schwartz
J.-M.
,
Weber
I.
,
Gerisch
G.
(
1997
).
Three-dimensional patterns and redistribution of myosin II and actin in mitotic Dictyostelium cells.
J. Cell Biol
139
,
1793
1804
Niewöhner
J.
,
Weber
I.
,
Maniak
M.
,
Muller-Taubenberger
A.
,
Gerisch.
G.
(
1997
).
Talin-null cells of Dictyostelium are strongly defective in adhesion to particle and substrate surfaces and slightly impaired in cytokinesis.
J. Cell Biol
138
,
349
361
Novak
K. D.
,
Peterson
M. D.
,
Reedy
M. C.
,
Titus
M. A.
(
1995
).
Dictyostelium myosin I double mutants exhibit conditional defects in pinocytosis.
J. Cell Biol
131
,
1205
1221
Raff
J. W.
,
Glover
D. M.
(
1989
).
Centrosomes, and not nuclei, initiate pole cell formation in Drosophila embryos.
Cell
57
,
611
619
Rappaport
R.
(
1961
).
Experiments concerning the cleavage stimulus in sand dollar eggs.
J. Exp. Zool
148
,
81
89
Rappaport
R.
(
1986
).
Establishment of the mechanism of cytokinesis in animal cells.
Intern. Rev. Cytol
105
,
245
281
Roos
U.-P.
,
Cattelan-Kohler
H.
(
1989
).
Formation and dynamics of the mitotic spindle in the cellular slime mold Polysphondylium violaceum.
Eur. J. Cell Biol
50
,
56
65
Shaw
S. L.
,
Yeh
E.
,
Maddox
P.
,
Salmon
E. D.
,
Bloom
K.
(
1997
).
Astral microtubule dynamics in yeast: a microtubule-based searching mechanism for spindle orientation and nuclear migration into the bud.
J. Cell Biol
139
,
985
994
Schejter
E. D.
,
Wieschaus
E.
(
1993
).
Functional elements of the cytoskeleton in the early Drosophila embryo.
Annu. Rev. Cell Biol
9
,
67
99
Schroeder
T. E.
(
1981
).
The origin of cleavage forces in dividing eggs. A mechanism in two steps.
Exp. Cell Res
134
,
231
240
Simson
R.
,
Wallraff
E.
,
Faix
J.
,
Niewöhner
J.
,
Gerisch
G.
,
Sackmann
E.
(
1998
).
Membrane bending modulus and adhesion energy of wild-type and mutant cells of Dictyostelium lacking talin or cortexillins.
Biophys. J
74
,
514
522
Spudich
J. A.
(
1989
).
In pursuit of myosin function.
Cell Regulation
1
,
1
11
Straight
A. F.
,
Marshall
W. F.
,
Sedat
J. W.
,
Murray
A. W.
(
1997
).
Mitosis in living budding yeast: anaphase A but no metaphase plate.
Science
277
,
574
578
Swann
M. M.
,
Mitchison
J. M.
(
1953
).
Cleavage of sea-urchin eggs in colchicine.
J. exp. Biol
30
,
506
513
Trivinos-Lagos
L.
,
Ohmachi
T.
,
Albrightson
C.
,
Burns
R. G.
,
Ennis
H. L.
,
Chisholm
R. L.
(
1993
).
The highly divergent-and -tubulins from Dictyostelium discoideum are encoded by single genes.
J. Cell Sci
105
,
903
911
Tuxworth
R. I.
,
Cheetham
J. L.
,
Machesky
L. M.
,
Spiegelmann
G. B.
,
Weeks
G.
,
Insall
R. H.
(
1997
).
Dictyostelium RasG is required for normal motility and cytokinesis, but not growth.
J. Cell Biol
138
,
605
614
Warrick
H. M.
,
Spudich
J. A.
(
1987
).
Myosin structure and function in cell motility.
Annu. Rev. Cell Biol
3
,
379
421
Wheatley
S. P.
,
Hinchcliffe
E. H.
,
Glotzer
M.
,
Hyman
A. A.
,
Sluter
G.
,
Wang
Y.-L.
(
1997
).
CDK1 inactivation regulates anaphase spindle dynamics and cytokinesis in vivo.
J. Cell Biol
138
,
385
393
White
J. G.
,
Borisy
G. G.
(
1983
).
On the mechanisms of cytokinesis in animal cells.
J. Theor. Biol
101
,
289
316
Wolpert
L.
(
1966
).
The mechanical properties of the membrane of the sea urchin egg during cleavage.
Exp. Cell Res
41
,
385
396
Yumura
S.
,
Mori
H.
,
Fukui
Y.
(
1984
).
Localization of actin and myosin for the study of ameboid movement in Dictyostelium using improved immunofluorescence.
J. Cell Biol
99
,
894
899
This content is only available via PDF.