The monoclonal antibody CTR210 raised against isolated human centrosomes strongly decorates the centrosome and more weakly a domain congruent with the Golgi apparatus in several animal cells (HeLa, 3T3, CHO, PtK2). Both decorations resist Triton extraction in conditions which totally extract the Golgi apparatus, as judged by galactosyltransferase decoration. A 67 kDa centrosomal antigen can be demonstrated in human cells with this antibody. CTR210 also decorates the centrosome or associated structures in several systems, including unicellular eukaryotes such as dinoflagellates or ciliates. A 72 kDa antigen has been identified and purified from the dinoflagellate C. cohnii and its NH2-terminal sequence partially established. It shows a close homology with HSP70 proteins. The possibility that the 72 kDa antigen belongs to this chaperone family was further supported using a mAb reacting, in most species, with HSP70. A polyclonal antibody raised against the 72 kDa antigen from C. cohnii decorates the centrosome in human cells and reacts with the CTR210 centrosomal 67 kDa antigen. These results suggest that specific chaperone proteins are associated with the centrosome in eukaryotic cells. The centrosomal chaperones could participate in the microtubule nucleation reaction or in the process of centrosome assembly.

REFERENCES

Ang
D.
,
Liberek
K.
,
Skowyra
D.
,
Zylicz
M.
,
Georgopoulos
C.
(
1991
).
Biological role and regulation of the universally conserved heat shock proteins.
J. Biol. Chem
266
,
24233
24236
Bailly
E.
,
Doree
M.
,
Nurse
P.
,
Bornens
M.
(
1989
).
p34cdc2is located in both nucleus and cytoplasm part is centrosomally associated at G2/M and enters vesicles at anaphase.
EMBO J
8
,
3985
3995
Bailly
E.
,
Pines
J.
,
Hunter
T.
,
Bornens
M.
(
1992
).
Cytoplasmic accumulation of cyclin B1 in human cells: association with a detergent-resistant compartment and with the centrosome.
J. Cell Sci
101
,
529
545
Baron
A. T.
,
Salisbury
J. L.
(
1988
).
Identification and localization of anovel cytoskeletal, centrosome-associated protein in PtK2cells.
J. Cell Biol
107
,
2669
2678
Barrau
M. D.
,
Blackburn
G. R.
,
Dewey
W. C.
(
1978
).
Effects of heat on the centrosomes of Chinese hamster ovary cells.
Cancer Res
38
,
2290
2294
Beckmann
R. P.
,
Mizzen
L. A.
,
Welch
W. J.
(
1990
).
Interactions of HSP70 with newly synthesized proteins: implications for protein folding and assembly.
Science
248
,
850
854
Berger
E. G.
,
Mandel
T.
,
Schilt
U.
(
1981
).
Immunhistochemical localization of galactosyltransferase a human fibroblasts and Hela cells.
J. Histochem. Cytochem
29
,
364
370
Bhaud
Y.
,
Salmon
J. M.
,
Soyer-Gobillard.
M. O.
(
1991
).
The complex cell cycle of the dinoflagellate protoctist Crypthecodinium cohnii as studied in vivo and by cytofluorimetry.
J. Cell Sci
100
,
675
682
Blisnick
T.
,
Lema
F.
,
Mazie
J. C.
,
Pereira da Silva
L.
(
1988
).
Plasmodium falciparum: analysis of B epitopes of a polypeptide antigen expressed in Escherichia coli, using monoclonal antibodies.
Exp. Parasitol
67
,
247
256
Bornens
M.
,
Paintrand
M.
,
Berges
J.
,
Marty
M. C.
,
Karsenti
E.
(
1987
).
Structural and chemical characterisation of isolated centrosomes.
Cell Motil. Cytoskel
8
,
238
249
Byers
B.
,
Goetsh.
L.
(
1975
).
Behavior of spindles and spindle plaques in the cell cycle and conjugation of Saccharomyces cerevisae.
J. Bacteriol
124
,
511
523
Cheng
M. Y.
,
Hartl
F. U.
,
Martin
J.
,
Pollock
R. A.
,
Kalousek
F.
,
Neupert
W.
,
Hallberg
E. M.
,
Hallberg
R. L.
,
Horwich
A. L.
(
1989
).
Mitochondrial heat-shock protein hsp60 is essential for assembly of proteins imported into yeast mitochondria.
Nature
337
,
620
625
Chirico
W. J.
,
Walters
M. J.
,
Blobel
G.
(
1988
).
70K heat shock related proteins stimulate protein translocation into microsomes.
Nature
332
,
805
809
Chitinis
P. R.
,
Nelson
N.
(
1991
).
Molecular cloning of the genes encoding two chaperone proteins of the cyanobacterium Synechocystis sp.
J. Biol. Chem
266
,
58
65
Clark
S. W.
,
Meyer
D. I.
(
1992
).
Centractin is an actin homologue associated with the centrosome.
Nature
359
,
246
250
Craig
E. A.
,
Kramer
J.
,
Shilling
J.
,
Werner-Washburne
M.
,
Holmes
S.
,
Kosic-Smithers
J.
,
Nicolet
C. M.
(
1989
).
SSC1 an esential member of the yeast HSP70 multigene family encodes a mitochondrial protein.
Mol. Cell. Biol
9
,
3000
3008
Debec
A.
,
Courgeon
A. M.
,
Maingourd
M.
,
Maisonhaute
C.
(
1990
).
The response of the centrosome to heat shock and related stresses in a Drosophila cell line.
J. Cell Sci
96
,
403
412
Deshaies
R. J.
,
Koch
B. D.
,
Werner-Washburne
M.
,
Craig
E. A.
,
Schekman
R.
(
1988
).
A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides.
Nature
332
,
800
805
Dessen
P.
,
Fondrat
C.
,
Valencien
C.
,
Mugnier
C.
(
1990
).
Bisance: a French service for access to biomolecular sequence databases.
Comp. Appl. Biosci
6
,
355
356
Devereux
J.
,
Haerberli
P.
,
Smithies
O.
(
1984
).
A comprehensive set of sequence analysis programs for the VAX.
Nucl. Acids Res
12
,
387
395
Dirksen
E. R.
(
1991
).
Centriole and basal body formation during ciliogenesis revisited.
Biol. Cell
72
,
31
38
Dress
V. M.
,
Yi
H.
,
Musal
M. R.
,
Williams
N. E.
(
1992
).
Tetrin polypeptides are colocalized in the cortex of Tetrahymena.
J. Struct. Biol
108
,
187
194
Ellis
R. J.
(
1987
).
Proteins as molecular chaperones.
Nature
328
,
378
379
Ellis
R. J.
,
Hemmigsen
S. M.
(
1989
).
Molecular chaperones: proteins essential for the biogenesis of some macromolecular structures.
Trends Biochem. Sci
14
,
339
342
Engman
D. M.
,
Kirchhoff
L. V.
,
Donelson
J. E.
(
1989
).
Molecular cloning of mtp70, a mitochondrial member of the hsp70 family.
Mol. Cell. Biol
9
,
5163
5168
Fleury
A.
,
Le Guyader
H.
,
Iftode
F.
,
Laurent
M.
,
Bornens.
M.
(
1993
).
A scaffold for basal body patterning revealed by a monoclonal antibody in the hypotrich ciliate Paraurostyla weissei.
Dev. Biol
157
,
285
302
Frydman
J.
,
Nimmesgern
E.
,
Erdjument-Bromage
H.
,
Wall
J. S.
,
Tempst
P.
,
Hartl.
F. U.
(
1992
).
Function un protein folding of TriC, acytosolic ring complex containing TCP-1 and structurally related subunits.
EMBO J
11
,
4767
4778
Gao
Y.
,
Thomas
J. O.
,
Chow
R. L.
,
Lee
G. H.
,
Cowan
N. J.
(
1992
).
A cytoplasmic chaperonin that catalyses-actin folding.
Cell
69
,
1043
1050
Gao
Y.
,
Vainberg
I. E.
,
Chow
R. L.
,
Lee
G. H.
,
Cowan
N. J.
(
1993
).
Two cofactors and cytoplasmic chaperonin are required for the folding ofand tubulin.
Mol. Cell Biol
13
,
2478
2485
Gething
M. J.
,
Sambrook
J.
(
1992
).
Protein folding in the cell.
Nature
355
,
33
45
Goloubinoff
P.
,
Christeller
J. T.
,
Gatenby
A. A.
,
Lorimer
G. H.
(
1989
).
Reconstitution of active dimeric ribulose bisphosphate carboxylase from an unfolded state depends on two chaperonin proteins and Mg-ATP.
Nature
342
,
884
889
Gosti
F.
,
Marty
M. C.
,
Berges
J.
,
Maunoury
R.
,
Bornens.
M.
(
1986
).
Identification of centrosomal proteins in a human lymphoblastic cell line.
EMBO J
5
,
2545
2550
Gundersen
G. G.
,
Bulinski.
J. C.
(
1986
).
Microtubule arrays in differentiated cells contain elevated levels of a post-transcriptionally modified form of tubulin.
Eur. J. Cell Biol
42
,
288
294
Gupta
R.
(
1990
).
Mitochondria, molecular chaperone proteins and the in vivo assembly of microtubules.
Trends Biochem. Sci
15
,
415
418
Hemmingsen
S. M.
,
Woolford
C.
,
van der Vies
S. M.
,
Tilly
K.
,
Dennis
D. T.
,
Georgopoulos
C. P.
,
Hendrix
R. W.
,
Ellis
R. J.
(
1988
).
Homologous plant and bacterial proteins chaperone oligomeric protein assembly.
Nature
333
,
330
334
Hendrix
R. W.
(
1979
).
Purification and properties of groE, a host protein involved in bacteriophage assembly.
J. Mol. Biol
129
,
375
392
Hohn
T.
,
Hohn
B.
,
Engel
A.
,
Wurtz
M.
,
Smith.
P. R.
(
1979
).
Isolation and characterization of the host protein groE involved in bacteriophage lambda assembly.
J. Mol. Biol
129
,
359
373
Huang
B.
,
Watterson
D. M.
,
Lee
V. D.
,
Schibler
M. J.
(
1988
).
Purification and characterization of a basal body-associated Ca2+-binding protein.
J. Cell Biol
107
,
121
131
Huang
B.
,
Mengersen
A.
,
Lee
V. D.
(
1988
).
Molecular cloning of cDNA for caltractin, a basal body-associated Ca2+-binding protein: homology in its protein sequence with calmodulin and yeast CDC31 gene product.
J. Cell Biol
107
,
133
140
Karsenti
E.
,
Newport
J.
,
Kirschner
M.
(
1984
).
Respective roles of centrosomes and chromatin in the conversion of microtubule arrays from interphase to metaphase.
J. Cell Biol
89
,
47
–.
Keryer
G.
,
Iftode
F.
,
Bornens
M.
(
1990
).
Identification of proteins associated with microtubule-organizing centres and filaments in the oral apparatus of the ciliate Paramecium tetraurelia.
J. Cell Sci
97
,
553
563
Klotz
C.
,
Dabauvale
M. C.
,
Paintrand
M.
,
Weber
T.
,
Bornens
M.
,
Karsenti.
E.
(
1990
).
Parthogenesis in Xenopus eggs requires centrosomal integrity.
J. Cell Biol
110
,
405
415
Kreis
T. E.
(
1990
).
Role of microtubules in the organisation of the Golgi apparatus.
Cell. Motil. Cytoskel
15
,
67
70
Laemmli
U. K.
(
1970
).
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature
227
,
680
685
Lewis
V. A.
,
Hynes
G. M.
,
Zheng
D.
,
Saibil
H.
,
Willison
K.
(
1992
).
T-complex polypeptide-1 is a subunit of a heteromeric particle in the eukaryotic cytosol.
Nature
358
,
249
252
Lim
L.
,
Hall
C.
,
Leung
T.
,
Whatley
S.
(
1984
).
The relationship of the rat brain 68 kDa microtubule-associated protein with synaptosomal plasma membranes and with the Drosophila 70 kDa heat shock protein.
Biochem. J
224
,
677
680
Lin
W.
,
Fung
B.
,
Shyamala
M.
,
Kasamatsu
H.
(
1981
).
Identification of antigenically related polypeptides at centrioles and basal bodies.
Proc. Nat. Acad. Sci.USA
78
,
2372
2377
Lingappa
J. R.
,
Martin
R. L.
,
Wong
M. L.
,
Ganem
D.
,
Welch
W. J.
,
Lingappa
V. R.
(
1994
).
A eukaryotic cytosolic chaperonin is associated with a high molecular weight intermediate in the assembly of Hepatitis B virus capsid a multimeric particle.
J. Cell Biol
125
,
99
111
Malawista
S. E.
,
De Boisfleury-Chevance
(
1982
).
The cytokineplast:purified, stable and functional motile machinery from human blood polymorphonuclear leukocytes.
J. Cell Biol
95
,
960
973
Maller
J.
,
Poccia
D.
,
Nishioka
D.
,
Kido
P.
,
Gerhart
J.
,
Hartman
H.
(
1976
).
Spindle formation and cleavage in Xenopus eggs injected with centriole containing fractions from sperm.
Exp. Cell Res
99
,
285
294
Marchesi
V. T.
,
Ngo.
N.
(
1993
).
In vitro assembly of multiprotein complexes containing, andtubulin, heat shock protein HSP70, and elongation factor 1.
Proc. Nat. Acad. Sci. USA
90
,
3028
3032
Mattei
D.
,
Scherf
A.
,
Bensaude
O.
,
DaSilva
L. P.
(
1989
).
A heat shock-like protein from the human malaria parasite Plasmodium falciparum induces autoantibodies.
Eur. J. Immunol
19
,
1823
1828
Mayer
L. F.
,
Shu Man
,
Kunkel.
H. G.
(
1982
).
Human T cell hybridoma secreting factors for IgA-specific help, polyclonal B cell activation, and B cell proliferation.
J. Exp. Med
156
,
1860
1865
Melki
R.
,
Vainberg
I. E.
,
Chow
R. L.
,
Cowan
N. J.
(
1993
).
Chaperonin-mediated folding of vertebrate actin-related protein and-tubulin.
J. Cell Biol
122
,
1301
1310
Melkonian
M.
,
Schulze
D.
,
McFadden
G. I.
,
Robenek.
H.
(
1988
).
A polyclonal antibody anticentrin distinguishes between two types of fibrous flagellar roots in green algae.
Protoplasma
144
,
56
61
Meyer
Y.
,
Grosset
J.
,
Chartier
Y.
,
Cleyet-Marel.
J. C.
(
1988
).
Preparation by two-dimensional electrophoresis of proteins for antibody production: antibodies against proteins whose synthesis is reduced by auxin in tobacco mesophyll protoplasts.
Electrophoresis
9
,
704
712
Moudjou
M.
,
Paintrand
M.
,
Vigues
B.
,
Bornens.
M.
(
1991
).
A human centrosomal protein is immunologically related to basal body-associated proteins from lower eucaryotes and is involved in the nucleation of microtubules.
J. Cell Biol
115
,
129
140
Neoh
S. H.
,
Gordon
C.
,
Potter
A.
,
Zola.
H.
(
1986
).
The purification of mouse monoclonal antibodies from ascitic fluid.
J. Immunol. Meth
91
,
231
235
Normington
K.
,
Kohno
K.
,
Kozutsumu
Y.
,
Gething
M. J.
,
Sambrook
J.
(
1989
).
S. cerevisae encodes an essential protein homologous in sequence and function to mammalian Bip.
Cell
57
,
1223
1236
Oakley
C. E.
,
Oakley.
B. R.
(
1989
).
Identification ofTubulin, a new member of the tubulin superfamily encoded by mipA gene of Aspergillus nidulans.
Nature
338
,
662
664
Oakley
B. R.
,
Oakley
C. E.
,
Yoon
Y.
,
Jung.
K.
(
1990
).
Tubulin is a component of the spindle pole body that is essential for microtubule function in Aspergillus nidulans.
Cell
61
,
1289
1301
Paintrand
M.
,
Moudjou
M.
,
Delacroix
H.
,
Bornens
M.
(
1992
).
Centrosome organization and centriole architecture: their sensitivity to divalent cations.
J. Struct. Biol
108
,
107
128
Perret
E.
,
Albert
M.
,
Bordes
N.
,
Bornens
M.
,
Soyer-Gobillard
M. O.
(
1991
).
Microtubular spindle and centrosome structures during cell cycle in a dinoflagellate Crypthecodinium cohnii B. an immunocytochemical study.
BioSystems
24
,
53
65
Perret
E.
,
Davoust
J.
,
Albert
M.
,
Besseau
L.
,
Soyer-Gobillard
M. O.
(
1993
).
Microtubular spindle and centrosome structures during the cell cycle in a Dinoflagellate Crypthecodinium cohnii B: an immunocytochemical study.
J. Cell Sci
104
,
639
651
Prevelige
P.
,
Thomas
D.
,
King.
J.
(
1988
).
Scaffolding protein regulates the polymerization of P22 coat subunits into icosahedral shells in vitro.
J. Mol. Biol
202
,
743
757
Rattner
J. B.
(
1990
).
HSP70 is localized to the centrosome of dividing HeLa cells.
Cell Motil. Cytoskel
17
,
227
235
Reading
D. S.
,
Halberg
R. L.
,
Myers
A. M.
(
1989
).
Characterization of the yeast HSP60 gene coding for a mitochondrial assembly factor.
Nature
337
,
655
659
Rothman
J. E.
(
1989
).
Polypeptide chain binding proteins: catalysts of protein folding and related processes in cells.
Cell
59
,
591
601
Rose
D. M.
,
Misra
L. M.
,
Vogel
J. P.
(
1989
).
Kar 2 a karyogamy gene is the yeast homolog of the mammalian Bip / GRP78 gene.
Cell
57
,
2111
2121
Rout
M. P.
,
Kilmartin
J. V.
(
1990
).
Components of the yeast spindle and spindle pole body.
J. Cell Biol
11
,
1913
1927
Salisbury
J. L.
,
Baron
A.
,
Surek
B.
,
Melkonian
M.
(
1984
).
Striated flagellar roots isolation and partial characterization of a calcium modulated contractile organelle.
J. Cell Biol
99
,
962
970
Salisbury
J. L.
,
Baron
A.
,
Sanders
M. A.
(
1988
).
The centrin-based cytoskeleton of Chlamydomonas reinhardtii: distribution in interphase and mitotic cells.
J. Cell Biol
107
,
635
641
Sigel
M.
,
Sinha
Y. N.
,
Van der Laan
W. P.
(
1983
).
Production of antibodies by inoculation into lymph nodes.
Meth. Enzymol
93
,
3
23
Stearns
T.
,
Evans
L.
,
Kirschner.
M.
(
1991
).
-Tubulin is a highly conserved component of the centrosome.
Cell
65
,
825
836
Sternlicht
H.
,
Farr
G. W.
,
Sternlicht
M. L.
,
Driscoll
J. K.
,
Willison
K.
,
Yaffe
M. B.
(
1993
).
The t-complex polypeptide 1 complex is a chaperonin for tubulin and actin in vivo.
Proc. Nat. Acad. Sci. USA
93
,
9422
9426
Tassin
A. M.
,
Paintrand
M.
,
Berger
E. G.
,
Bornens
M.
(
1985
).
The Golgi apparatus remains asociated with microtubule organizing centers during myogenesis.
J. Cell Biol
101
,
630
638
Ting
J.
,
Lee.
A. S.
(
1988
).
Human gene encoding the 78. 000 dalton glucose regulated protein and its pseudogene: structure conservation and regulation.
DNA
7
,
275
286
Tokuyasu
K. T.
(
1989
).
Use of poly(vinylpyrrolidone) and poly(vinylalcohol) for cryoultramicrotomy.
Histochem. J
21
,
163
171
Tournier
F.
,
Karsenti
E.
,
Bornens.
M.
(
1989
).
Parthenogenesis in Xenopus eggs injected with centrosomes from synchronized human lymphoid cells.
Dev. Biol
136
,
321
329
Tournier
F.
,
Cyrklaff
M.
,
Karsenti
E.
,
Bornens
M.
(
1991
).
Centrosomes competent for parthenogenesis in Xenopus eggs support procentriole budding in cell free extracts.
Proc. Nat. Acad. Sci. USA
88
,
9929
9933
Towbin
H.
,
Staehelin
T.
,
Gordon
J.
(
1979
).
Electrophoretic transfer of proteins from acrylamide gels to nitrocellulose sheets: procedure and some applications.
Proc. Nat. Acad. Sci.USA
76
,
4350
4354
Trent
J. T.
,
Nimmesgern
E.
,
Wall
J. S.
,
Hartl
F. U.
,
Horwich
A. L.
(
1991
).
A molecular chaperone from a thermophilic archaebacterium is related to the eukaryotic protein t-complex polypeptide 1.
Nature
354
,
490
493
Tuttle
R. C.
,
Loeblich.
A. R.
(
1975
).
An optimal growth medium for Dinoflagellate Crypthecodinium cohnii.
Phycologia
14
,
1
8
Ursic
D.
,
Culbertson.
M. R.
(
1991
).
The yeast homolog to mouse Tcp 1 affects microtubule mediated process.
Mol. Cell. Biol
11
,
2629
2640
Vidair
C. A.
,
Doxsey
S. J.
,
Dewey.
W. C.
(
1993
).
Heat shock alters centrosome organization leading to mitotic dysfunction and cell death.
J. Cell. Physiol
154
,
443
455
Weller
N.
(
1988
).
A 70 kDa microtubule-associated protein in NIL8 cells comigrates with the 70 kDa heat shock protein.
Biol. Cell
63
,
307
317
Werner-Washburne
M.
,
Stone
D. E.
,
Craig
E. A.
(
1987
).
Complex interactions among members of an essential subfamilly of hsp70 genes in Saccharomyces cerevisiae.
Mol. Cell. Biol
7
,
2568
2577
Yaffe
M. B.
,
Farr
G. W.
,
Miklos
D.
,
Horwih
A. L.
,
Sternlicht
M. L.
,
Sternlicht
H.
(
1992
).
TCP 1 complex is a molecular chaperone in tubulin biogenesis.
Nature
358
,
248
248
Zheng
Y.
,
Jung
M. K.
,
Oakley
B. R.
(
1991
).
-Tubulin is present in Drosophila melanogaster and Homo sapiens and is associated with the centrosome.
Cell
65
,
817
823
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