The Escherichia coli cell division protein FtsZ was expressed in Chinese hamster ovary cells, where it formed a striking array of dots that were independent of the mammalian cytoskeleton. Although FtsZ appears to be a bacterial homolog of tubulin, its expression had no detectable effects on the microtubule network or cell growth. However, treatment of the cells with vinblastine at concentrations that caused microtubule disassembly rapidly induced a network of FtsZ filaments that grew from and connected the dots, suggesting that the dots are an active storage form of FtsZ. Cells producing FtsZ also exhibited vinblastine- and calcium-resistant tubulin polymers that colocalized with the FtsZ network. The FtsZ polymers could be selectively disassembled, indicating that the two proteins were not copolymerized. The vinblastine effects were readily reversible by washing out the drug or by treating the cells with the vinblastine competitor, maytansine. These results demonstrate that FtsZ assembly can occur in the absence of bacterial chaperones or cofactors, that FtsZ and tubulin do not copolymerize, and that tubulin-vinblastine complexes have an enhanced ability to interact with FtsZ.
REFERENCES
REFERENCES
Barlow
S. B.
, Gonzalez-Garay
M. L.
, West
R. R.
, Olmsted
J. B.
, Cabral
F.
(1994
). Stable expression of heterologous microtubule associated proteins in Chinese hamster ovary cells: evidence for differingroles of MAPs in microtubule organization.
J. Cell Biol
126
, 1017
–1029
Bensch
K. G.
, Malawista
S. E.
(1969
). Microtubular crystals in mammalian cells.
J. Cell Biol
40
, 95
–106
Bhattacharyya
B.
, Wolff
J.
(1977
). Maytansine binding to the vinblastine sites of tubulin.
FEBS Lett
75
, 159
–162
Boggs
B.
, Cabral
F.
(1987
). Mutations affecting assembly and stability of tubulin: evidence for a non-essential-tubulin in CHO cells.
Mol. Cell. Biol
7
, 2700
–2707
Bramhill
D.
(1997
). Bacterial cell division.
Annu. Rev. Cell Dev. Biol
13
, 395
–424
Cabral
F.
, Schatz
G.
(1979
). One-and two-dimensional electrophoretic analysis of mitochondrial membrane proteins.
Methods Enzymol
56
, 602
–613
Casaregola
S.
, Chen
M.
, Bouquin
N.
, Norris
V.
, Jacq
A.
, Goldberg
M.
, Margarson
S.
, Tempete
M.
, McKenna
S.
, Sweetman
H.
, Bernard
S.
, McGurk
G.
, Seror
S.
, Holland
I. B.
(1991
). Analysis of a myosin-like protein and the role of calcium in the E. coli cell cycle.
Res. Microbiol
142
, 201
–207
Cormack
B. P.
, Valdivia
R. H.
, Falkow
S.
(1996
). FACS-optimized mutants of the green fluorescent protein (GFP).
Gene
173
, 33
–38
de Boer
P.
, Crossley
R.
, Rothfield
L.
(1992
). The essential bacterial cell-division protein FtsZ is a GTPase.
Nature
359
, 254
–256
Erickson
H. P.
, Taylor
D. W.
, Taylor
K. A.
, Bramhill
D.
(1996
). Bacterial cell division protein FtsZ assembles into protofilament sheets and minirings, structural homologs of tubulin polymers.
Proc. Natl. Acad. Sci. USA
93
, 519
–523
Faguy
D. M.
, Doolittle
W. F.
(1998
). Cytoskeletal proteins: the evolution of cell division.
Curr. Biol
8
, 338
–341
Gonzalez-Garay
M. L.
, Cabral
F.
(1995
). Overexpression of an epitope-tagged-tubulin in Chinese hamster ovary cells causes an increase in endogenous -tubulin synthesis.
Cell Motil. Cytoskel
31
, 259
–272
Gonzalez-Garay
M. L.
, Cabral
F.
(1996
). -tubulin limits its own synthesis: evidence for a mechanism involving translational repression.
J. Cell Biol
135
, 1525
–1534
Gossen
M.
, Bujard
H.
(1992
). Tight control of gene expression in mammalian cells by tetracycline-responsive promoters.
Proc. Natl. Acad. Sci. USA
89
, 5547
–5551
Hiller
G.
, Weber
K.
(1978
). Radioimmunoassay for tubulin: a quantitative comparison of the tubulin content of different established tissue culture cells and tissues.
Cell
14
, 795
–804
Liang
P.
, MacRae
T. H.
(1997
). Molecular chaperones and the cytoskeleton.
J. Cell Sci
110
, 1431
–1440
Löwe
J.
, Amos
L. A.
(1998
). Crystal structure of the bacterial cell-division protein FtsZ.
Nature
391
, 203
–.Lu
C.
, Stricker
J.
, Erickson
H. P.
(1998
). FtsZ from Escherichia coli, Azotobacter vinelandii, and Thermotoga maritima —quantitation, GTP hydrolysis, and assembly.
Cell Motil. Cytoskel
40
, 71
–86
Lutkenhaus
J.
, Addinall
S. G.
(1997
). Bacterial cell division and the Z ring.
Ann. Rev. Biochem
66
, 93
–116
Ma
X.
, Ehrhardt
D. W.
, Margolin
W.
(1996
). Colocalization of cell division proteins FtsZ and FtsA to cytoskeletal structures in living Escherichia coli cells by using green fluorescent protein.
Proc. Natl. Acad. Sci. USA
93
, 12998
–13003
Malawista
S. E.
, Sato
H.
(1969
). Vinblastine produces uniaxial, birefringent crystals in starfish oocytes.
J. Cell Biol
42
, 596
–599
Margolin
W.
(1998
). A green light for the bacterial cytoskeleton.
Trends Microbiol
6
, 233
–238
Margolin
W.
, Wang
R.
, Kumar
M.
(1996
). Isolation of an ftsZ homolog from the archaebacterium Halobacterium salinarium: implications for the evolution of FtsZ and tubulin.
J. Bacteriol
178
, 1320
–1327
Minotti
A. M.
, Barlow
S. B.
, Cabral
F.
(1991
). Resistance to antimitotic drugs in Chinese hamster ovary cells correlates with changes in the level of polymerized tubulin.
J. Biol. Chem
266
, 3987
–3994
Mukherjee
A.
, Dai
K.
, Lutkenhaus
J.
(1993
). Escherichia coli cell division protein FtsZ is a guanine nucleotide binding protein.
Proc. Natl. Acad. Sci. USA
90
, 1053
–1057
Mukherjee
A.
, Lutkenhaus
J.
(1998
). Dynamic assembly of FtsZ regulated by GTP hydrolysis.
EMBO J
17
, 462
–469
Nogales
E.
, Wolf
S. G.
, Downing
K. H.
(1998
). Structure of the alpha-beta tubulin dimer by electron crystallography.
Nature
391
, 199
–203
Osteryoung
K. W.
, Stokes
K. D.
, Rutherford
S. M.
, Percival
A. L.
, Lee
W. Y.
(1998
). Chloroplast division in higher plants requires members of two functionally divergent gene families with homology to bacterial ftsZ.
Plant Cell
10
, 1991
–2004
Raychaudhuri
D.
, Park
J. T.
(1992
). Escherichia coli cell-division gene ftsZ encodes a novel GTP-binding protein.
Nature
359
, 251
–254
Rothfield
L. I.
, Justice
S. S.
(1997
). Bacterial cell division: the cycle of the ring.
Cell
88
, 581
–584
Sackett
D. L.
(1995
). Vinca site agents induce structural changes in tubulin different from and antagonistic to changes induced by colchicine site agents.
Biochemistry
34
, 7010
–7019
Sawada
T.
, Cabral
F.
(1989
). Expression and function of-tubulin isotypes in Chinese hamster ovary cells.
J. Biol. Chem
264
, 3013
–3020
Spiegelman
B. M.
, Penningroth
S. M.
, Kirschner
M. W.
(1977
). Turnover of tubulin and the N site GTP in Chinese hamster ovary cells.
Cell
12
, 587
–600
Strepp
R.
, Scholz
S.
, Kruse
S.
, Speth
V.
, Reski
R.
(1998
). Plant nuclear gene knockout reveals a role in plastid division for the homolog of the bacterial cell division protein FtsZ, an ancestral tubulin.
Proc. Natl. Acad. Sci. USA
95
, 4368
–4373
Sun
Q.
, Yu
X.-C.
, Margolin
W.
(1998
). Assembly of the FtsZ ring at the central division site in the absence of the chromosome.
Mol. Microbiol
29
, 491
–504
Tian
G.
, Huang
Y.
, Rommelaere
H.
, Vandekerckhove
J.
, Ampe
C.
, Cowan
N. J.
(1996
). Pathway leading to correctly folded beta-tubulin.
Cell
86
, 287
–296
Wang
X.
, Lutkenhaus
J.
(1996
). FtsZ ring: the eubacterial division apparatus conserved in archaebacteria.
Mol. Microbiol
21
, 313
–320
Weisenberg
R. C.
(1972
). Microtubule formation in vitro in solutions containing low calcium concentrations.
Science
177
, 1104
–1105
Yu
X.-C.
, Margolin
W.
(1997
). Ca2+-mediated GTP-dependent dynamic assembly of bacterial cell division protein FtsZ into asters and polymer networks in vitro.
EMBO J
16
, 5455
–5463
Yu
X.-C.
, Margolin
W.
(1998
). Inhibition of assembly of bacterial cell division protein FtsZ by the hydrophobic dye bis-8-anilino-1-naphthalenesulfonate.
J. Biol. Chem
273
, 10216
–10222
Yu
X.-C.
, Tran
A. H.
, Sun
Q.
, Margolin
W.
(1998
). Localization of cell division protein FtsK to the Escherichia coli septum and identification of a potential N-terminal targeting domain.
J. Bacteriol
180
, 1296
–1304
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