The BimC family of kinesin like proteins are involved in spindle dynamics in a wide variety of organisms. The human member of this family, HsEg5, has been implicated in centrosome separation during prophase/prometaphase and in the organization of in vitro mitotic asters. HsEg5 displays a complex distribution during mitosis, associating with the centrosomes, spindle microtubules, specific regions of the intracellular bridge and a microtubule bundle that forms in association with the post-mitotic migration of the centrosome. In an effort to determine the function of HsEg5 during late mitotic events and refine its proposed function during early mitotic centrosome separation, we microinjected antibodies specific to HsEg5 into HeLa cells during various stages of mitosis. In the presence of HsEg5 antibodies we find that the microtubule arrays responsible for both pre- and post-mitotic centrosome movement never form. Similarly, the microtubule bundle within the intracellular bridge becomes prematurely altered following karyokinesis resulting in the loss of the microtubule array at either end of the bridge. In addition, some peri-centrosomal material at the spindle poles becomes fragmented and the distribution of the spindle protein NuMA becomes more concentrated at the minus ends of the spindle microtubules. Our study also provides direct evidence that there is a link between post-mitotic centrosome migration and Golgi complex positioning and reformation following mitosis. We conclude that HsEg5 plays a recurrent role in establishing and/or determining the stability of specific microtubule arrays that form during cell division and that this role may encompass the ability of HsEg5 to influence the distribution of other protein components associated with cell division

REFERENCES

Ault
J. G.
,
Rieder
C. L.
(
1994
).
Centrosome and kinetochore movement during mitosis.
Curr. Opin. Cell Biol
6
,
41
49
Blangy
A.
,
Lane
H. A.
,
Herin
P.
,
Harper
M.
,
Kress
M.
,
Nigg
E. A.
(
1995
).
Phosphorylation by p34cdc2regulates spindle association of human Eg5, a kinesin-related motor protein essential for bipolar spindle formation in vivo.
Cell
83
,
1159
1169
Blangy
A.
,
Arnaud
L.
,
Nigg
E. A.
(
1997
).
Phosphorylation by p34cdc2protein kinase regulates binding of the kinesin-related motor HsEg5 to the dynactin subunit p150.
J. Biol. Chem
272
,
19418
19424
Blangy
A.
,
Chaussepied
P.
,
Nigg
E. A.
(
1998
).
Rigor-type mutation in the kinesin-related protein HsEg5 changes its subcellular localization and induces microtubule bundling.
Cell Motil. Cytoskel
40
,
174
182
Compton
D. A.
,
Yen
T. J.
,
Cleveland
D. W.
(
1991
).
Identification of novel centromere/kinetochore-associated proteins using monoclonal antibodies generated against human mitotic chromosome scaffolds.
J. Cell Biol
112
,
1083
1097
Corthesy-Theulaz
I.
,
Pauloin
A.
,
Pfeffer
R.
(
1992
).
Cytoplasmic dynein participates in the centrosomal localization of the Golgi complex.
J. Cell Biol
118
,
1333
1345
Duden
R.
,
Griffiths
G.
,
Frank
R.
,
Argos
P.
,
Kreis
T. E.
(
1991
).
-COP, a 110 kd protein associated with non-clathrin-coated vesicles and the Golgi complex, shows homology to -adaptin.
Cell
64
,
649
665
Feramisco
J. R.
(
1979
).
Microinjection of fluorescently labeled alpha-actinin into living fibroblasts.
Proc. Nat. Acad. Sci. USA
76
,
3967
3971
Gaglio
T.
,
Dionne
M. A.
,
Compton
D. A.
(
1997
).
Mitotic spindle poles are organized by structural and motor proteins in addition to centrosomes.
J. Cell Biol
138
,
1055
1066
Gaglio
T.
,
Saredi
A.
,
Bingham
J. B.
,
Hasbani
M. J.
,
Gill
S. R.
,
Schroer
T. A.
,
Compton
D. A.
(
1997
).
Opposing motor activities are required for the organization of the mammalian mitotic spindle pole.
J. Cell Biol
135
,
399
414
Griffith
K. J.
,
Chan
E. K.
,
Lung
C.
,
Hamel
J. C.
,
Guo
X.
,
Miyachi
K.
,
Fritzler
M. J.
(
1997
).
Molecular cloning of a novel 97-kd Golgi complex autoantigen associated with Sjögren's Syndrome.
Arthritis Rheum
40
,
1693
1702
Hagan
I.
,
Yanagida
M.
(
1992
).
Kinesin-related cut7 protein associates with mitotic and meiotic spindles in fission yeast.
Nature
356
,
74
76
Ho
W. C.
,
Allan
V. J.
,
Meer
G.
,
Berger
E. G.
,
Kreis
T. E.
(
1989
).
Reclustering of scattered Golgi elements occurs along microtubules.
Eur. J. Cell Biol
48
,
250
263
LeGueliec
R.
,
Paric
J.
,
Couturier
A.
,
Roghi
C.
,
Philippe
M.
(
1991
).
Cloning by differential screening of a Xenopus cDNA that encodes a kinesin-related protein.
Mol. Cell. Biol
11
,
3395
3398
Lucocq
J. M.
,
Pryde
J. G.
,
Berger
E. G.
,
arren
G.
(
1987
).
A mitotic form of the Golgi apparatus in HeLa cells.
J. Cell Biol
104
,
865
874
Lucocq
J. M.
,
Berger
E. G.
,
Warren
G.
(
1989
).
Mitotic Golgi fragments in HeLa cells and their role in the reassembly pathway.
J. Cell Biol
109
,
463
474
Mack
G.
,
Rattner
J. B.
(
1993
).
Centrosome repositioning immediately following karyokinesis and prior to cytokinesis.
Cell Motil. Cytoskel
26
,
239
247
Mack
G. J.
,
Rees
J.
,
Sandblom
O.
,
Balczon
R.
,
Fritzler
M. J.
,
Rattner
J. B.
(
1998
).
Autoantibodies to a group of centrosomal proteins in human autoimmune sera reactive with the centrosome.
Arthritis Rheum
41
,
551
558
Moskalewski
S.
,
Popowicz
P.
,
Tyberg
J.
(
1994
).
Functions of the Golgi complex in cell division: formation of cell-matrix contact and cell-cell communication channels in the terminal phase of cytokinesis.
J. Submicrosc. Cytol. Pathol
26
,
9
20
Orci
L.
,
Tagaya
M.
,
Amherdt
M.
,
Perrelet
A.
,
Donaldson
J. A.
,
Lippincott-Schwartz
J.
,
Klausner
R. D.
,
Rothman
J. E.
(
1991
).
Brefeldin A, a drug that blocks secretion, prevents the assembly of non-clathrin-coated buds on Golgi cisternae.
Cell
64
,
1183
1195
Rattner
J. B.
,
Wang
T.
,
Mack
G.
,
Martin
L.
,
Fritzler
M. J.
(
1992
).
MSA-35: a protein identified by human autoantibodies that colocalizes with microtubules.
Biochem. Cell Biol
70
,
1115
1122
Saunders
W. S.
,
Koshland
D.
,
Eshel
D.
,
Gibbons
I. R.
,
Hoyt
M. A.
(
1995
).
Saccharomyces cerevisiae kinesin-and dynein-related proteinsrequired for anaphase chromosome segregation.
J. Cell Biol
128
,
617
624
Sawin
K. E.
,
LeGuellec
K.
,
Philippe
M.
,
Mitchison
T. J.
(
1992
).
Mitotic spindle organization by a plus-end-directed microtubule motor.
Nature
359
,
540
543
Sawin
K. E.
,
Mitchison
T. J.
(
1995
).
Mutations in the kinesin-like protein Eg5 disrupting localization to the mitotic spindle.
Proc. Nat. Acad. Sci. USA
92
,
4289
4293
Thyberg
J.
,
Moskalewski
S.
(
1992
).
Reorganization of the Golgi complex in association with mitosis: redistribution of mannosidase II to the endoplasmic reticulum and effects of brefeldin A.
J. Submicrosc. Cytol. Pathol
24
,
495
508
Walczak
C. E.
,
Mitchison
T. J.
(
1996
).
Kinesin-related proteins at mitotic spindle poles: Function and regulation.
Cell
85
,
943
946
Waters
J. C.
,
Cole
R. W.
,
Rieder
C. L.
(
1993
).
The force-producing mechanism for centrosome separation during spindle formation in vertebrates is intrinsic to each aster.
J. Cell Biol
122
,
361
372
Whitehead
C. M.
,
Winkfein
R. J.
,
Fritzler
M. J.
,
Rattner
J. B.
(
1996
).
The spindle kinesin-like protein HsEg5 is an autoantigen in systemic lupus erythematosus.
Arthritis Rheum
39
,
1635
1642
Whitehead
C. M.
,
Winkfein
R. J.
,
Rattner
J. B.
(
1996
).
The relationship of HsEg5 and the actin cytoskeleton to centrosome separation.
Cell Motil. Cytoskel
35
,
298
308
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