Using a new subtraction method and chemically induced rat hepatocellular carcinomas, we identified a hepatocellular carcinogenesis and hepatocyte proliferation-related gene designated hp33 that encoded a 33-kDa protein. The predicted protein was similar to the bovine aralkyl N-acyltransferase and arylacetyl N-acyltransferase. HP33 was restrictively expressed in the liver and kidney, and its gene expression was stimulated in the regenerating liver as well as in hepatocellular carcinoma. Interestingly, it was demonstrated in various hepatic cells that HP33 was localized in regions surrounding the centrosome, where mitochondria were not concentrated. Moreover, its centrosomal localization was evident in the interphase but not in the mitotic phase of the cell cycle. The centrosomal localization of HP33 was dependent on microtubules, and ectopically expressed HP33 was seen at centrosomes even in fibroblasts, which do not exhibit a typical staining pattern of HP33. The centrosomal localization of HP33 became invisible by nocodazole treatment, whereas the mitochondrial staining pattern was not affected by it. In vitro cosedimentation experiments using purified microtubules indicated that HP33 bound to MTs directly and that its MT-binding ability was dependent on the C-terminal basic domain of the protein. These results suggest that, different from early predictions based on its primary structure, HP33 has a growth- and carcinogenesis-related function that may be independent of mitochondrial function.

Bardelli
A.
,
Pugliese
L.
,
Comoglio
P. M.
(
1997
).
‘Invasive-growth’ signaling by the Met/HGF receptor: the hereditary renal carcinoma connection.
Biochim. Biophys. Acta
1333
,
41
–.
Fiszer-Szafarz
B.
,
Nadal
C.
(
1977
).
Lysosomal enzyme activities in the regenerating rat liver.
Cancer Res
37
,
354
357
Francavilla
A.
,
Vujanovic
N. L.
,
Polimeno
L.
,
Azzarone
A.
,
Lacobellis
A.
,
Deleo
A.
,
Hagiya
M.
,
Whiteside
T. L.
,
Starzl
T. E.
(
1997
).
The in vivo effect of hepatotrophic factors augmenter of liver regeneration, hepatocyte growth factor, and insulin-like growth factor-II on liver natural killer cell functions.
Hepatology
25
,
411
415
Funabiki
H.
,
Yamano
H.
,
Kumada
K.
,
Nagao
K.
,
Hunt
T.
,
Yanagida
M.
(
1996
).
Cut2 proteolysis required for sister-chromatid seperation in fission yeast.
Nature
381
,
438
441
Gonzalez
C.
,
Tavosanis
G.
,
Mollinari
C.
(
1998
).
Centrosomes and microtubule organisation during Drosophila development.
J. Cell Sci
111
,
2697
2706
Gubler
U.
,
Hoffman
B. J.
(
1983
).
A simple and very efficient method for generating cDNA libraries.
Gene
2
,
263
269
Hanna
D. E.
,
Rethinaswamy
A.
,
Glover
C. V.
(
1995
).
Casein kinase II is required for cell cycle progression during G1and G2/M in Saccharomyces cerevisiae.
J. Biol. Chem
270
,
25905
25914
Igawa
T.
,
Kanda
S.
,
Kanetake
H.
,
Saitoh
Y.
,
Ichihara
A.
,
Tomita
Y.
,
Nakamura
T.
(
1991
).
Hepatocyte growth factor is a potent mitogen for cultured rabbit renal tubular epithelial cells.
Biochem. Biophys. Res. Commun
174
,
831
838
Irminger-Finger
I.
,
Laymon
R. A.
,
Goldstein
L. S.
(
1990
).
Analysis of the primary sequence and microtubule-binding region of the Drosophila 205K MAP.
J. Cell Biol
111
,
2563
2572
Irniger
S.
,
Piatti
S.
,
Michaelis
C.
,
Nasmyth
K.
(
1995
).
Genes involved in sister chromatid separation are needed for B-type cyclin proteolysis in budding yeast.
Cell
81
,
269
278
Johnson
M.
,
Koukoulis
G.
,
Matsumoto
K.
,
Nakamura
T.
,
Iyer
A.
(
1993
).
Hepatocyte growth factor induces proliferation and morphogenesis in nonparenchymal epithelial liver cells.
Hepatology
17
,
1052
1061
Kellogg
D. R.
,
Moritz
M.
,
Alberts
B. M.
(
1994
).
The centrosome and cellular organization.
Annu. Rev. Biochem
63
,
639
674
Kinoshita
T.
,
Hirao
S.
,
Matsumoto
K.
,
Nakamura
T.
(
1991
).
Possible endocrine control by hepatocyte growth factor of liver regeneration after partial hepatectomy.
Biochem. Biophys. Res. Commun
31
,
330
335
Kishimoto
T.
,
Kokura
K.
,
Nakadai
T.
,
Miyazawa
Y.
,
Wakamatsu
T.
,
Makino
Y.
,
Nakamura
T.
,
Hara
E.
,
Oda
K.
,
Muramatsu
M.
,
Tamura
T.
(
1996
).
Overexpression of cysteine sulfinic acid decarboxylase stimulated by hepatocarcinogenesis results in autoantibody production in rats.
Cancer Res
56
,
5230
5237
Kishimoto
T.
,
Kokura
K.
,
Ohkawa
N.
,
Makino
Y.
,
Yoshida
M.
,
Hirohashi
S.
,
Niwa
S.
,
Muramatsu
M.
,
Tamura
T.
(
1998
).
Enhanced expression of a new class of liver-enriched b-Zip transcription factors, hepatocarcinogenesis-related transcription factor, in hepatocellular carcinomas of rats and humans.
Cell Growth Differ
9
,
337
344
Kobayashi
N.
,
Mundel
P.
(
1998
).
A role of microtubules during the formation of cell processes in neuronal and non-neuronal cells.
Cell Tissue Res
291
,
163
174
Kloc
M.
,
Etkin
L. D.
(
1995
).
Two distinct pathways for the localization of RNAs at the vegetal cortex in Xenopus oocytes.
Development
121
,
287
297
Krek
W.
,
Maridor
G.
,
Nigg
E. A.
(
1992
).
Casein kinase II is a predominantly nuclear enzyme.
J. Cell Biol
116
,
43
55
Krendel
M.
,
Sgourdas
G.
,
Bonder
E. M.
(
1998
).
Disassembly of actin filaments leads to increased rate and frequency of mitochondrial movement along microtubules.
Cell Motil. Cytoskel
40
,
368
378
Kuriyama
R.
,
Nislow
C.
(
1992
).
Molecular components of the mitotic spindle.
BioEssays
14
,
81
88
Langford
G. M.
(
1995
).
Actin-and microtubule-dependent organelle motors: interrelationships between the two motility systems.
Curr. Opin. Cell Biol
7
,
82
88
Lisowsky
T.
(
1996
).
Removal of an intron with unique 3branch site creates an amino-terminal protein sequence directing the scERV1 gene product to mitochondria.
Yeast
15
,
1501
1510
Li
P.
,
Nijhawan
D.
,
Budihardjo
I.
,
Srinivasula
S. M.
,
Ahmad
M.
,
Alnemri
E. S.
,
Wang
X.
(
1997
).
Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade.
Cell
91
,
479
489
Maekawa
T.
,
Leslie
R.
,
Kuriyama
R.
(
1991
).
Identification of a minus end-specific microtubule-associated protein located at the mitotic poles in cultured mammalian cells.
Eur. J. Cell Biol
54
,
255
67
Matsumoto
K.
,
Nakamura
T.
(
1992
).
Hepatocyte growth factor: molecular structure, roles in liver regeneration, and other biological functions.
Crit. Rev. Oncog
3
,
27
54
Merdes
A.
,
Ramyar
K.
,
Vechio
J. D.
,
Cleveland
D. W.
(
1996
).
A complex of NuMA and cytoplasmic dynein is essential for mitotic spindle assembly.
Cell
87
,
447
458
Montesano
R.
,
Matsumoto
K.
,
Nakamura
T.
,
Orci
L.
(
1991
).
Identification of a fibroblast-derived epithelial morphogen as hepatocyte growth factor.
Cell
67
,
901
908
Murray
N. R.
,
Burns
D. J.
,
Fields
A. P.
(
1994
).
Presence of a beta II protein kinase C-selective nuclear membrane activation factor in human leukemia cells.
J. Biol. Chem
269
,
21385
21390
Nakadai
T.
,
Kishimoto
T.
,
Kokura
K.
,
Ohkawa
N.
,
Makino
Y.
,
Muramatsu
M.
,
Tamura
T.
(
1998
).
Cloning of a novel rat gene, DB83, that encodes a putative membrane protein.
DNA Res
5
,
1
3
Nigg
E. A.
,
Schafer
G.
,
Hilz
H.
,
Eppenberger
H. M.
(
1985
).
Cyclic-AMP-dependent protein kinase type II is associated with the Golgi complex and with centrosomes.
Cell
41
,
1039
1051
Pereira
G.
,
Schiebel
E.
(
1997
).
Centrosome-microtubule nucleation.
J. Cell Sci
110
,
295
300
Raff
J. W.
,
Kellogg
D. R.
,
Alberts
B. M.
(
1993
).
Drosophila gamma-tubulin is part of a complex containing two previously identified centrosomal MAPs.
J. Cell Biol
121
,
823
835
Ramadori
G.
,
Neubauer
K.
,
Odenthal
M.
,
Nakamura
T.
,
Knittel
T.
,
Schwogler
S.
,
Meyer zum Buschenfelde
K. H.
(
1992
).
The gene of hepatocyte growth factor is expressed in fat-storing cells of rat liver and is downregulated during cell growth and by transforming growth factor-beta.
Biochem. Biophys. Res. Commun
183
,
739
742
Solt
F.
,
Farber
E.
(
1976
).
New principle for the analysis of chemical carcinogenesis.
Nature
263
,
701
703
Studier
F. W.
,
Rosenberg
A. H.
,
Dunn
J. J.
,
Dubendorff
J. W.
(
1990
).
Use of T7 RNA polymerase to direct expression of cloned genes.
Methods Enzymol
185
,
60
89
Tajima
H.
,
Matsumoto
K.
,
Nakamura
T.
(
1991
).
Hepatocyte growth factor has potent anti-proliferative activity in various tumor cell lines.
FEBS Lett
291
,
229
232
Takahashi
M.
,
Ota
S.
,
Terano
A.
,
Yoshiura
K.
,
Matsumura
M.
,
Niwa
Y.
,
Kawabe
T.
,
Nakamura
T.
,
Omata
M.
(
1993
).
Hepatocyte growth factor induces mitogenic reaction to the rabbit gastric epithelial cells in primary culture.
Biochem. Biophys. Res. Commun
191
,
528
534
Vessey
D.
,
Lau
E.
(
1996
).
Determination of the sequence of the aralkyl acyl-CoA: amino acid N-acyltransferase from bovine liver mitochondria.
J. Biochem. Toxicol
11
,
211
215
Wells
N. J.
,
Fry
A. M.
,
Guano
F.
,
Norbury
C.
,
Hickson
I. D.
(
1995
).
Cell cycle phase-specific phosphorylation of human topoisomerase II alpha. Evidence of a role for protein kinase C.
J. Biol. Chem
270
,
28357
28363
Yamada
M.
,
Okigaki
T.
,
Awai
M.
(
1987
).
Role of superoxide radicals in cytotoxic effects of Fe-NTA on cultured normal liver epithelial cells.
Cell Struct. Funct
12
,
407
420
Zorov
D. B.
,
Krasnikov
B. F.
,
Kuzminova
A. E.
,
Vysokikh
M. Y.
,
Zorova
L. D.
(
1997
).
Mitochondria revisited. Alternative functions of mitochondria.
Biosci. Rep
17
,
507
520
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