We have identified, by differential cDNA library screening, 15 serum inducible genes in the human diploid fibroblast cell line WI-38. The genes fall into two classes that are distinguished by their dependence on protein synthesis for the induction by serum, i.e., primary and secondary genes. While 11 of these genes encode known proteins, 4 other genes have not been described to date. The former genes encode proteins of diverse functions, including the monocyte-derived neutrophil chemotactic factor (MONAP), calmodulin, tropomyosin, tenascin, collagenase, plasminogen activator inhibitor-2a, the ‘sperm-specific’ cleavage signal-1 protein, metallothionein IIa and the mitochondrial chaperonin hsp-60. Interestingly, one of the unknown genes contains a large open reading frame for a polypeptide that is highly homologous to a previously unidentified long open reading frame in the opposite strand of the gene coding for the transcription factor HTF-4. We also studied the regulation of these serum-induced genes during cell cycle progression in normally cycling WI-38 and HL-60 cells separated by counterflow elutriation as well as in serum-stimulated HL-60 cells. Our results clearly show that, in contrast to the prevailing opinion, the expression of most genes induced after mitogen stimulation is not subject to a significant regulation in normally proliferating cells. This supports the hypothesis that the progression into S from either G0 or G1 are distinct processes with specific patterns of gene expression.

REFERENCES

Almendral
J. M.
,
Sommer
D.
,
Macdonald-Bravo
H.
,
Burckhardt
J.
,
Perera
J.
,
Bravo
R.
(
1988
).
Complexity of the early genetic response to growth factors in mouse fibroblasts.
Mol. Cell. Biol
8
,
2140
2148
Angel
P.
,
Baumann
I.
,
Stein
B.
,
Delius
H.
,
Rahmsdorf
H. J.
,
Herrlich
P.
(
1987
).
12-O-Tetradecanoyl-phorbol-13-acetate induction of the human collagenase gene is mediated by an inducible enhancer element located in the 5-flanking region.
Mol. Cell. Biol
7
,
2256
2266
Angel
P.
,
Imagawa
M.
,
Chiu
R.
,
Stein
B.
,
Imbra
R. J.
,
Rahmsdorf
H. H.
,
Jonat
C.
,
Herrlich
P.
,
Karin
M.
(
1987
).
Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor.
Cell
49
,
729
739
Armelin
H. A.
,
Armelin
M. C. S.
,
Kelly
K.
,
Steward
T.
,
Leder
P.
,
Cochran
B. H.
,
Stiles
C. D.
(
1984
).
Functional role of c- myc in mitogenic response to platelet-derived growth factor.
Nature
310
,
655
660
Borus
V.
,
Villalobos
J.
,
Burd
P. R.
,
Kelly
K.
,
Siebenlist
U.
(
1990
).
Cloning of a mitogen-inducible gene encoding aB DNA-binding protein with homology to the rel oncogene and to cell-cycle motifs.
Nature
348
,
76
80
Bravo
R.
(
1990
).
Growth factor-responsive genes in fibroblasts.
Cell Growth Diff
1
,
305
309
Bravo
R.
,
Burckhardt
J.
,
Curran
T.
,
Muller
R.
(
1986
).
Expression of c- fos in NIH 3T3 cells is very low but inducible throughout the cell cycle.
EMBO J
5
,
695
700
Burger
C.
,
Wick
M.
,
Brusselbach
S.
,
Muller
R.
(
1994
).
Differential induction of ‘metabolic genes’ after mitogen stimulation and during normal cell cycle progression.
J. Cell Sci
107
,
241
252
Calabretta
B.
,
Battini
R.
,
Kaczmarek
L.
,
DeReil
J. K.
,
Baserga
R.
(
1986
).
Molecular cloning of the cDNA for a growth factor-inducible gene with strong homology to S-100, a calcium-binding protein.
J. Biol. Chem
261
,
12628
12632
Chafouleas
J. G.
,
Lagace
L.
,
Bolton
W. E.
,
Boyd
A. E.
III
,
Means
A.R.
) (
1984
).
Changes in Calmodulin and its mRNA accompany reentry of quiescent(G0) cells into the cell cycle.
Cell
36
,
73
81
Chomczynski
P.
,
Sacchi
N.
(
1987
).
Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
Anal. Biochem
162
,
156
159
Cochran
B. H.
,
Reffel
A. C.
,
Stiles
C. D.
(
1983
).
Molecular cloning of gene sequences regulated by platelet-derived growth factor.
Cell
33
,
939
947
Coppock
D. L.
,
Pardee
A. B.
(
1985
).
Regulation of thymidine kinase mRNA during the mammalian cell cycle (abstr.).
J. Cell Biol
101
,
79
–.
Dalton
S.
(
1992
).
Cell cycle regulation of the human cdc2 gene.
EMBO J
11
,
1797
1804
Greenberg
M. E.
,
Ziff
E. B.
(
1984
).
Stimulation of3T3 cells induces transcription of the c- fos proto-oncogene.
Nature
311
,
433
438
Hann
S. R.
,
Thompson
C. B.
,
Eisenman
R. N.
(
1985
).
c- myc oncogene protein synthesis is independent of the cell cycle in human and avian cells.
Nature
314
,
366
369
Hayflick
L.
(
1965
).
The limited in vitro lifetime of human diploid cell strains.
Exp. Cell Res
37
,
614
636
Hazel
T. G.
,
Nathans
D.
,
Lau
L. F.
(
1988
).
A gene inducible by serum growth factors encodes a member of the steroid and thyroid hormone receptor superfamily.
Proc. Nat. Acad. Sci. USA
85
,
8444
8448
Heikkila
R.
,
Schwab
G.
,
Wickstrom
E.
,
Loke
S. L.
,
Pluznik
D. H.
,
Watt
R.
,
Neckers
L. M.
(
1987
).
A c- myc antisense oligodeoxynucleotide inhibits entry into S phase but not progress from G0to G1.
Nature
328
,
445
449
Hershman
H. R.
(
1991
).
Primary response genes induced by growth factors and tumor promoters.
Annu. Rev. Biochem
60
,
281
320
Herzog
H.
,
Hoefferer
L.
,
Schneider
R.
,
Schweiger
M.
(
1990
).
cDNA encoding the human homologue of rat ribosomal protein L35a.
Nucl. Acids Res
18
,
4600
–.
Hiraki
Y.
,
Rosen
O. M.
,
Birnbaum
M. J.
(
1988
).
Growth factors rapidly induce expression of the glucose transporter gene.
J. Biol. Chem
263
,
13655
13662
Javed
A. A.
,
Naz
R. K.
(
1992
).
Human cleavage signal-1 protein; cDNA cloning, transcription and immunological analysis.
Gene
112
,
205
211
Johnson
R. S.
,
Spiegelmann
B. M.
,
Papaioannou
V.
(
1992
).
Pleiotropic effects of a null mutation in c-fos proto-oncogene.
Cell
71
,
577
–.
Jones
F. S.
,
Crossin
K. L.
,
Cunningham
B. A.
,
Edelman
G. M.
(
1990
).
Identification and characterization of the promoter for the cytoactin gene.
Proc. Nat. Acad. Sci. USA
87
,
6497
6501
Kaczmarek
L.
,
Hyland
J. K.
,
Watt
R.
,
Rosenberg
M.
,
Baserga
R.
(
1985
).
Microinjected c- myc as a competence factor.
Science
228
,
1313
1315
Karin
M.
,
Haslinger
A.
,
Holtgreve
H.
,
Richards
R. I.
,
Krauter
P.
,
Westphal
H. M.
,
Beato
M.
(
1984
).
Characterization of DNA sequences through which cadmium and glucocorticoid hormones induce human metallothionein IIa gene.
Nature
308
,
513
519
Kelly
K.
,
Cochran
B. H.
,
Stiles
C. D.
,
Leder
P.
(
1983
).
Cell-specific regulation of the c- myc gene by lymphocyte mitogens and platelet-derived growth factor.
Cell
35
,
603
610
Kho
C.-J.
,
Zarbl
H.
(
1992
).
Fte-1, a v- fos transformation effector gene, encodes the mammalian homologue of a yeast gene involved in protein import into mitochondria.
Proc. Natl. Acad. Sci. USA
89
,
2200
2204
Kovary
K.
,
Bravo
R.
(
1991
).
The jun and fos protein families are both required for cell cycle progression in fibroblasts.
Mol. Cell. Biol
11
,
4466
4472
Kovary
K.
,
Bravo
R.
(
1992
).
Existence of different Fos/Jun complexes during the G0-to-G1transition and during exponential growth in mouse fibroblasts: differential role of Fos proteins.
Mol. Cell. Biol
12
,
5015
5023
Kowalski
J.
,
Denhardt
D. T.
(
1989
).
Regulation of the mRNA for monocyte-derived neutrophil-activating peptide in differentiating HL-60 promyelocytes.
Mol. Cell. Biol
9
,
1946
1957
Lanahan
A.
,
Williams
J. B.
,
Sanders
L. K.
,
Nathans
D.
(
1992
).
Growth factor-induced delayed early response genes.
Mol. Cell. Biol
12
,
3919
3929
Lau
L. F.
,
Nathans
D.
(
1985
).
Identification of a set of genes expressed during the G0/G1transition of cultured mouse cells.
EMBO J
4
,
3145
3151
Lee
M. G.
,
Nurse
P.
(
1987
).
Complementation used to clone a humanhomologue of the fission yeast cell cycle control gene cdc2.
Nature
327
,
31
35
Lee
W.
,
Haslinger
A.
,
Karin
M.
,
Tjian
R.
(
1987
).
Activation of transcription by two factors that bind promoter and enhancer sequences of the human metallothionein gene and SV40.
Nature
324
,
368
372
Lin
C.-S.
,
Leavitt
J.
(
1988
).
Cloning and characterization of a cDNA encoding transformation sensitive tropomyosin isoform 3 from tumorigenic human fibroblasts.
Mol. Cell. Biol
8
,
160
168
Mackie
E. J.
,
Halfter
W.
,
Liverani
D.
(
1988
).
Induction of tenascin in healing wounds.
J. Cell Biol
107
,
2757
2767
Masibay
A. S.
,
Qasba
P. K.
,
Sengupta
D. N.
,
Damewood
G. P.
,
Sreevalsan
T.
(
1988
).
Cell-cycle-specific and serum-dependent expression of gamma-actin mRNA in Swiss mouse 3T3 cells.
Mol. Cell. Biol
8
,
2288
2294
McDonnell
S. E.
,
Kerr
L. D.
,
Matrisian
L. M.
(
1990
).
Epidermal growth factor stimulation of stromelysin mRNA in rat fibroblasts requires induction of proto-oncogenes c-fos and c-jun and activation of protein kinase C.
Mol. Cell. Biol
10
,
4284
4293
Milbrandt
J.
(
1988
).
Nerve growth factor induces a gene homologous to the glucocorticoid receptor gene.
Neuron
1
,
183
188
Muller
R.
,
Slamon
D. J.
,
Adamson
E. D.
,
Tremblay
J. M.
,
Muller
D.
,
Cline
M. J.
,
Verma
I. M.
(
1983
).
Transcription of c- onc genes c- ras Kiand c- fms during mouse development.
Mol. Cell. Biol
3
,
1062
1069
Muller
R.
,
Bravo
R.
,
Burckhardt
J.
,
Curran
T.
(
1984
).
Induction of c- fos gene and protein by growth factors precedes activation of c- myc.
Nature
312
,
716
720
Muller
R.
,
Mumberg
D.
,
Lucibello
F. C.
(
1993
).
Signals and genes in cell cycle progression.
Biochim. Biophys. Acta
1155
,
151
179
Oquendo
P.
,
Alberta
J.
,
Wen
D. Z.
,
Graycar
J. L.
,
Derynck
R.
,
Stiles
C. D.
(
1989
).
The platelet-derived growth factor-inducible KC gene encodes a secretory protein related to platelet alpha-granule proteins.
J. Biol. Chem
264
,
4133
4137
Ponte
P.
,
Ng
S.-Y.
,
Engel
J.
,
Gunning
P.
,
Kedes
L.
(
1984
).
Evolutionary conservation in the untranslated regions of actin mRNAs: DNA sequence of a human beta-actin cDNA.
Nucl. Acids Res
12
,
1687
1696
Prochownik
E. V.
,
Kukowska
J.
,
Rodgers
C.
(
1988
).
c- myc antisense transcripts accelerate differentiation and inhibit G1progression in murine erythroleukemia cells.
Mol. Cell. Biol
8
,
3683
3695
Rasmussen
C. D.
,
Means
A. R.
(
1989
).
Calmodulin, cell growth, and gene expression.
Trends Neurosci
12
,
433
438
Riabowol
K.
,
Schiff
J.
,
Gilman
M. Z.
(
1992
).
Transcription factor AP-1 activity is required for initiation of DNA synthesis and is lost during cellular aging.
Proc. Nat. Acad. Sci. USA
89
,
157
161
Rittling
S. R.
,
Brooks
K. M.
,
Cristofalo
V. J.
,
Baserga
R.
(
1986
).
Expression of cell cycle-specific genes in young and senescent WI-38 fibroblasts.
Proc. Nat. Acad. Sci. USA
83
,
3316
3320
Ryder
K.
,
Nathans
D.
(
1988
).
Induction of protooncogene c- jun by serum growth factors.
Proc. Nat. Acad. Sci. USA
85
,
8464
8467
Ryder
K.
,
Lau
L. F.
,
Nathans
D.
(
1988
).
A gene activated by growth factors is related to the oncogene v- jun.
Proc. Nat. Acad. Sci. USA
85
,
1487
1491
Ryseck
R.-P.
,
Hirai
S. I.
,
Yaniv
M.
,
Bravo
R.
(
1988
).
Transcriptional activation of c- jun during the G0/G1transition in mouse fibroblasts.
Nature
334
,
535
537
Ryseck
R. P.
,
Mac Donald-Bravo
H.
,
Zerial
M.
,
Bravo
R.
(
1989
).
Coordinate induction of fibronectin, fibronectin receptor, tropomyosin, and actin gene in serum-stimulated fibroblasts.
Exp. Cell Res
180
,
537
545
SenGupta
B.
,
Friedberg
F.
,
Detera-Wadleigh
S. D.
(
1987
).
Molecular analysis of human and rat calmodulin complementary DNA clones. Evidence for additional active genes in these species.
J. Biol. Chem
262
,
16663
16670
Sewing
A.
,
Burger
C.
,
Brusselbach
S.
,
Schalk
C.
,
Lucibello
F. C.
,
Muller
R.
(
1993
).
Human cyclin D1 encodes a labile nuclear protein whose synthesis is directly induced by growth factors and suppressed by cyclic AMP.
J. Cell Sci
104
,
545
555
Siri
A.
,
Carnemolla
B.
,
Saginati
M.
,
Leprini
A.
,
Casari
G.
,
Baralle
P.
,
Zardi
L.
(
1991
).
Human tenascin: primary structure, pre-mRNA, splicing patterns and localization of the epitopes recognized by two monoclonal antibodies.
Nucl. Acids Res
19
,
525
531
Thompson
C. B.
,
Challoner
P. B.
,
Neiman
P. E.
,
Groudine
M.
(
1985
).
Levels of c- myc oncogene mRNA are invariant throughout the cell cycle.
Nature
314
,
363
366
Tirone
F.
,
Shooter
E. M.
(
1989
).
Early gene regulation by nerve growth factor in PC12 cells: Induction of an interferon-related gene.
Proc. Nat. Acad. Sci. USA
86
,
2088
2092
Venner
T. J.
,
Singh
B.
,
Gupta
R. S.
(
1990
).
Nucleotide sequences and novel structural features of human and Chinese hamster hsp60 (chaperonin) gene families.
DNA Cell Biol
9
,
545
552
Vincent
S.
,
Marty
L.
,
Le Gallic
L.
,
Jeanteur
P.
,
Fort
P.
(
1993
).
Characterization of late response genes sequentially expressed during renewed growth of fibroblastic cells.
Oncogene
8
,
1603
1610
Wahl
A. F.
,
Geis
A. M.
,
Spain
B. H.
,
Wong
S. W.
,
Korn
D.
,
Wang
T. S. F.
(
1988
).
Gene expression of human DNA polymerase alpha during cell proliferation and the cell cycle.
Mol. Cell. Biol
8
,
5016
5025
Wang
Z.-Q.
,
Ovitt
C.
,
Grigoriadis
A. E.
,
Möhle-Steinlein
U.
,
Ruther
U.
,
Wagner
E. F.
(
1992
).
Bone and haematopoietic defects in mice lacking c- fos.
Nature
360
,
741
745
Whitham
S. E.
,
Murphy
G.
,
Angel
P.
,
Rahmsdorf
H. J.
,
Smith
B. J.
,
Lyons
A.
,
Harris
T. J.
,
Reynolds
J. J.
,
Herrlich
P.
,
Docherty
A. J.
(
1986
).
Comparison of human stromelysin and collagenase by cloning and sequence analysis.
Biochem. J
240
,
913
916
Wickstrom
E. L.
,
Bacon
T. A.
,
Gonzalez
A.
,
Freeman
D. L.
,
Lyman
G. M.
,
Wickstrom
E.
(
1988
).
Human promelocytic leukemia HL-60 cell proliferation and c-myc protein synthesis are inhibited by an antisense pentadecadeoxynucleotide targeted against c-myc mRNA.
Proc. Nat. Acad. Sci. USA
85
,
1028
1032
Won
K. A.
,
Xiong
Y.
,
Beach
D.
,
Gilman
M. Z.
(
1992
).
Growth-regulated expression of D-type cyclin genes in human diploid fibroblasts.
Proc. Nat. Acad. Sci. USA
89
,
9910
–.
Yang
G. P.
,
Lau
L. F.
(
1991
).
Cyr61, product of a growth factor-inducible immediate-early gene, is associated with the extracellular matrix and the cell surface.
Cell Growth Differ
2
,
351
357
Yang
Q.
,
Tonk
N. K.
(
1991
).
Isolation of a cDNA clone encoding a human protein-tyrosine phosphatase with homology to the cytoskeletal-associated proteins band 4. 1, ezrin, and talin.
Proc. Nat. Acad. Sci. USA
88
,
5949
5953
Zhang
Y.
,
Babin
J.
,
Feldhaus
A. L.
,
Singh
H.
,
Sharp
P. A.
,
Bina
M.
(
1991
).
HTF: a new human helix-loop-helix protein.
Nucl. Acids Res
19
,
4555
–.
Zipfel
P. F.
,
Irving
S. G.
,
Kelly
K.
,
Siebenlist
U.
(
1989
).
Complexity of the primary genetic response to mitogenic activation of human T cells.
Mol. Cell. Biol
9
,
1041
1047
This content is only available via PDF.