From the Department of Gene Regulation, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan
Received for publication, October 23, 2000
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ABSTRACT |
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Fos family proteins form stable heterodimers with
Jun family proteins, and each heterodimer shows distinctive
transactivating potential for regulating cellular growth,
differentiation, and development via AP-1 binding sites. However, the
molecular mechanism underlying dimer specificity and the molecules that
facilitate transactivation remain undefined. Here, we show that BAF60a,
a subunit of the SWI·SNF chromatin remodeling complex, is a
determinant of the transactivation potential of Fos/Jun dimers. BAF60a
binds to a specific subset of Fos/Jun heterodimers using two different interfaces for c-Fos and c-Jun, respectively. Only when the functional SWI·SNF complex is present, can c-Fos/c-Jun (high affinity to BAF60a)
but not Fra-2/JunD (no affinity to BAF60a) induce the endogenous
AP-1-regulated genes such as collagenase and c-met. These results indicate that a specific subset of Fos/Jun dimers recruits SWI·SNF complex via BAF60a to initiate transcription.
Transcription factor AP-1, which plays pivotal roles in cell
growth, differentiation, development, and tumor formation, is composed
of Fos family proteins (Fos; c-Fos, Fra-1, Fra-2, and FosB) and Jun
family proteins (Jun; c-Jun, JunB, and JunD). The members of both
families form dimers through a leucine zipper structure; Jun
family members can form low-affinity homodimers and high affinity
heterodimers with the Fos family (1, 2). However, members of the Fos
family do not form stable homodimers. Although these hetero- and
homodimers bind to similar sites on DNA (TGAC/GTCA, AP-1 binding sites)
through the basic domains of both proteins, each dimer has a distinct
transcriptional regulatory function that can be modulated either
positively or negatively (3). For example, transcriptional activity of
the c-Fos/c-Jun dimer is much higher than the Fra-2/c-Jun dimer.
Although functional interactions between some members of the Fos/Jun
family of proteins and adaptor proteins such as CREB-binding
protein (CBP) or TATA-binding protein (TBP) have been reported (4, 5),
crucial proteins that recognize the dimer specificity and/or facilitate
the induction of transcriptional initiation were largely unknown.
Therefore, we proposed here to isolate such a crucial protein using a
yeast two-hybrid system and have demonstrated that BAF60a (6), a component of the SWI·SNF chromatin remodeling complex, can select specific Fos/Jun dimers and function as a determinant of
transcriptional activation via AP-1 binding sites.
Plasmid Construction--
The Gal4DBD-c-Jun () fusion
construct (pDBLeu-cJ(25-187)) was generated by inserting the 0.49 kilobase pair EheI fragment of the rat
c-jun gene into the MscI restriction endonuclease
cleavage site within the open reading frame of Gal4DBD. For the
construction of template DNA for in vitro translation, we
first generated a starter plasmid from pGEM2-475/Jun-D, the
translation initiation site of which has a Kozak consensus sequence
with a unique NcoI site that is preceded by a fragment 475 sequence for an efficient translational initiation (2). The
1.0-kilobase pair NcoI-BamHI fragment of
pGEM2-475/Jun-D encoding JunD was excised and substituted with a
synthetic DNA fragment carrying multiple cloning sites to generate
pGEM2-475/Met. A series of c-jun or c-fos
deletions was inserted in-frame into pGEM2-475/Met. Full-length BAF60a
cDNA was constructed by inserting a
PCR1-generated DNA fragment
encoding the N-terminal region into the original partial BAF60a clone,
which encodes BAF60a, amino acids 139-475, and was isolated by yeast
two-hybrid screening. Glutathione S-transferase (GST)-BAF60a
was constructed by inserting the full-length DNA into the pGEX4T vector
(Amersham Pharmacia Biotech). For the construction of a retrovirus
vector expressing HA-tagged proteins, a synthetic oligo DNA fragment
coding the HA sequence was inserted into the unique BamHI
site of pBabe-IRES puro (7) to generate pBabeHA-IRES puro. The DNA
fragment was generated by annealing the following two nucleotides:
5'-gatcctaccatgtatccatatgatgttccagattatgctagcctcgcctcgagtggccgacaagcgtctcgcgacggtataccgtgagtaagtagg-3' and
5'-gatccctacttactcacggtataccgtcgcgagacgcttgtcggccactcgaggcgaggctagcataatctggaacatcatatggatacatggtag-3'.
Yeast Two-hybrid Screening--
The yeast strain MaV203
(MAT Cell Culture, Transfection, and Virus
Transduction--
Virus-packaging cell line BOSC23 and the
adenocarcinoma cell line SW13 were grown in Dulbecco's modified
Eagle's medium supplemented with 10% fetal calf serum. NIH3T3 cells
were grown in Dulbecco's modified Eagle's medium supplemented with
10% calf serum. BOSC23 cells were transfected with pBabeHA-BAF60a-IRES
puro, and NIH3T3 cells were infected with the resulting ecotropic
retrovirus as described previously (8).
Protein Analysis--
The GST pull-down assay,
SDS-polyacrylamide gel electrophoresis, immunoprecipitation, and
Western blotting were performed as described previously (9) using
anti-Brm (Transduction Laboratories, Lexington, KY), anti-HA (Santa
Cruz Laboratories, Santa Cruz, CA), anti-c-Jun (Santa Cruz), anti-c-Fos
(Oncogene Science, San Diego, CA), and anti-INI1 (Santa Cruz)
antibodies. A gel shift assay was performed as described previously
(3).
Reverse Transcription-Polymerase Chain Reaction--
Total RNA
was prepared from SW13 cells transfected with expression plasmids using
an Isogen RNA isolation reagent (Wako) and reverse-transcribed for 30 min at 50 °C. Each PCR regime involved an initial denaturation step
at 94 °C, 3 min followed by 30 cycles (for collagenase and
c-met) and 25 cycles (for GAPDH) at 94 °C for 30 s,
56 °C for 1 min, and 72 °C for 1 min. RT-PCR was performed within the linear range with Superscript one-step RT-PCR with Platinum
Taq kit (Life Technologies, Inc.). The primer sets and amplified fragment size for RT-PCR were as follows: collagenase, 570 base pairs; forward 5'-atcttttgtcaggggagatcatcg-3', reverse 5'-acagcccagtacttattccctttg-3', c-met, 701 base pairs;
forward 5'-atgagcactgctttaataggacac-3', reverse
5'-accaactgtgcatttcaatgtattc-3', GAPDH, 431 base pairs; forward
5'-tcattgacctcaactacatggtttac-3', reverse
5'-ggcatggactgtggtcatgagtc-3'.
To identify the molecules involved in the transactivation of AP-1,
we first proposed to isolate proteins that specifically bind with rat
c-Jun N-terminal amino acids 25-187 (Fig.
1A), which reportedly contain
transactivation domains (10). Using the yeast two-hybrid system, 23 positive clones were obtained upon screening 5 × 105
clones of a mouse brain cDNA library. DNA sequence analysis
revealed a single cDNA clone encoding the C-terminal half of mouse
BAF60a, which is a subunit of the mouse SWI·SNF complex. The
SWI·SNF complex is a 2-MDa, multi-subunit, DNA-dependent
ATPase that is thought to disrupt repressive chromatin structure (11).
Because interaction of c-Jun with the SWI·SNF complex seemed likely
to be important for regulation of transcription in vivo, we
isolated a cDNA containing the entire open reading frame of BAF60a,
which had the same nucleotide sequence as a clone reported previously
(6). Escherichia coli-produced, histidine-tagged c-Jun amino
acids 1-225 (9) bound with GST-BAF60a fusion protein but not with GST
(data not shown), demonstrating that BAF60a interacts directly with
c-Jun. By synthesizing truncation mutants of c-Jun in reticulocyte
lysates, we showed that N-terminal amino acids 25-187 of c-Jun were
sufficient for binding, but further truncation at either the N-terminal
or C-terminal regions totally abolished binding (Fig.
1A).
INTRODUCTION
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
EXPERIMENTAL PROCEDURES
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
, leu2-3
112, trp1-901,
his3
200, ade2-101, gal4
,
gal80
, SPAL10:: URA3,
GAL1::lacZ, HIS
3uasGAL1::HIS3@Lys2, can1R, cyh2R)
harboring pDBLeu-cJ () (Fig. 1A) was transformed by
the lithium acetate method with a mouse brain cDNA fusion library
inserted into the activation domain vector, pPC86 (Life Technologies,
Inc.). Transformants were selected with a HIS3 reporter containing the Gal4 DNA binding sites by seeding onto SC/
Leu/
Trp/
His/+3AT (50 mM) plates. After incubation for 60 h at
30 °C, the plates were replica-cleaned for the reduction of
false-positive clones according to the manufacturer's protocol and
further incubated at 30 °C for 44 h. The positive clones were
confirmed by another promoter system that induces
-galactosidase expression through the same DNA binding
domain.
-Galactosidase activity in yeast was assayed according to
the manufacturer' s protocol.
RESULTS
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
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Fig. 1.
Binding properties of wild-type and deletion
mutants of c-Jun and c-Fos, and wild-type of their family
proteins. A, interaction between c-Jun N terminus and
BAF60a. Binding of a series of c-Jun truncation mutants that were
synthesized in reticulocyte lysates containing
[35S]methionine and GST-BAF60a was assessed by a GST
precipitation assay. The relative binding activities are designated
by plus and minus signs and represent the percentages of total
protein that precipitated: +++, >16%; ++, 16 4%; +, 4
0.05%; and
, <0.05%. The upper rectangle represents the
structure of the bait, which contains pDBLeu-cJ () used for the
yeast two-hybrid analysis, and the lower rectangle
represents the full-length rat c-Jun. The striped regions
represent the GAL4 DNA binding domain. B, interaction
between truncation mutants of rat c-Fos and BAF60a. The solid
bar indicates the frameshift region of v-Fos. The interaction
between GST-BAF60a and members of the Jun (C) or Fos
(D) families is shown. Wild-type c-Jun (cJ), JunB
(JB), or JunD (JD) (Jun family) or c-Fos
(cF), Fra-1 (F1), Fra-2 (F2), or FosB
(FB) (Fos family) were labeled with
[35S]methionine by in vitro translation in
reticulocyte lysates and incubated with purified GST-BAF60a attached to
GSH-Sepharose beads. The bound proteins were boiled in SDS and
separated by SDS-10% polyacrylamide gel electrophoresis. A fraction
(12.5%) of the total labeled Jun or Fos proteins was also analyzed on
the same gel (left side).
To examine whether BAF60a binds with other members of the Jun family, JunB and JunD proteins were synthesized in reticulocyte lysates and assayed for binding with a recombinant GST-BAF60a fusion protein. In comparison with BAF60a binding with c-Jun, BAF60a displayed weaker binding with JunB and no binding with JunD (Fig. 1C). This result was somewhat surprising because the BAF60a binding domain of c-Jun includes regions conserved in both JunB and JunD (conserved regions 1-3; Fig. 1A). Even more surprising was the finding that GST-BAF60a fusion protein bound with in vitro translated, full-length c-Fos (Fig. 1D); c-Fos and c-Jun share little sequence similarity outside of their leucine zipper domains. Among the Fos family proteins, this binding is specific for c-Fos (Fig. 1D). Analysis of c-Fos truncation mutants indicated that C-terminal amino acids 237-380 are sufficient for BAF60a binding (Fig. 1B). Binding of BAF60a with v-Fos, which is derived from FBJ-MuSV (12), was not detected (Fig. 1B). The C-terminal amino acids of v-Fos and c-Fos are divergent because of a frameshift mutation starting at c-Fos amino acid 333 (13) (Fig. 1B). Therefore, c-Fos C-terminal amino acids 333-380 are likely to be critical for binding with BAF60a.
Fos/Jun heterodimers composed of c-Fos and c-Jun, c-Fos and JunD, Fra-2
and c-Jun, or Fra-2 and JunD were assayed for binding with GST-BAF60a
by precipitation with glutathione (GSH)-Sepharose beads.
GST-BAF60a precipitated 4% of c-Fos alone. Precipitation of c-Fos with
BAF60a increased to 7.5% of the total c-Fos when preincubated with an
equimolar amount of c-Jun (Fig.
2A), Because the molar amount
of c-Jun recovered in the precipitate was nearly equal to the amount of
c-Fos, it seems that binding of BAF60a with the c-Fos/c-Jun heterodimer
would be preferential to binding with c-Fos alone. Fra-2 alone,
JunD alone, and the Fra-2/JunD heterodimer displayed only marginal
binding with BAF60a (Fig. 2D). JunD alone displayed marginal
binding with GST-BAF60a, whereas precipitation of the c-Fos/JunD
heterodimer with GST-BAF60a resulted in the recovery of 1.5% of the
total JunD added to the reaction mixture.
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These results suggest that BAF60a can simultaneously bind restricted members of Fos family proteins (c-Fos alone) and Jun family proteins (c-Jun, and probably to a lesser extent JunB but not JunD) using different interfaces present in the BAF60a molecule. These results also suggest that the Fos/Jun dimer formation via the leucine zipper region is compatible with formation of a ternary complex composed of Fos, Jun, and BAF60a. A c-Fos/JunD heterodimer bound with GST-BAF60a, but the binding was less than that observed with c-Fos alone (Fig. 2B), suggesting that dimerization of c-Fos and JunD did not appreciably contribute to BAF60a binding. Similarly, dimerization of Fra-2 and c-Jun did not seem to contribute to BAF60a binding (Fig. 2C). The binding affinity of each heterodimer examined here correlated positively with their transactivating activity when determined in F9 cells by transient expression with a reporter gene containing a single AP-1 binding site derived from the human collagenase gene (Table I). c-Fos/c-Jun had the highest transactivating activity, and other dimers consisting of either c-Fos or c-Jun had lower activity. Dimers that contained neither c-Fos nor c-Jun had only marginal activity.
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To examine whether BAF60a affects the specific DNA binding activity of
c-Fos/c-Jun heterodimer, electrophoretic mobility shift assays were
performed with 32P-labeled probe containing the AP-1 DNA
binding site of the human collagenase gene. Gel mobility shift was not
affected upon addition of GST alone, but a dramatic enhancement of the
shifted band was detected upon the addition of GST-BAF60a (Fig.
3A). These results indicate
that BAF60a stimulates the specific DNA binding activity of the
c-Fos/c-Jun heterodimer. When the Fra-2/JunD dimer was assayed, the
addition of GST-BAF60a caused only a slight enhancement of the shifted
band (data not shown).
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To study whether c-Fos/c-Jun dimer associates with BAF60a in vivo, NIH3T3 cells expressing HA-tagged BAF60a were growth-stimulated (G1) to induce endogeneous c-Fos and c-Jun. Cell lysates were mixed with a Ni2+ beads bound with histidine-tagged c-Jun. Proteins that bound with histidine-tagged-c-Jun were recovered and analyzed by Western blotting. They contained not only HA-tagged BAF60a but also Brm (14), the catalytic subunit (DNA-dependent ATPase) of the SWI·SNF complex (Fig. 3B). When cell lysates of growth-stimulated (G1) NIH3T3 cells were precipitated with anti-c-Jun antiserum and analyzed by Western blotting, the immunoprecipitates contained Brm and SWI·SNF complex subunit INI1 (Fig. 3C) (14). Both proteins disappeared when the anti-c-Jun antiserum was preabsorbed by the antigen. Similar results were obtained when immunoprecipitations were performed with anti-c-Fos antiserum. No Brm or INI1 proteins were detected when experiments were performed in growth-arrested (Go) cells, which express no Fos protein (Fig. 3C). Taken together, it was shown that c-Fos and c-Jun associate with the SWI·SNF complex through BAF60a in vivo.
These results indicate that BAF60a has a versatile protein interface
for c-Fos, c-Jun, and the other SWI·SNF complex subunits. Mutational
analysis (Fig. 4) of GST-BAF60a revealed
the location of a large domain spanning amino acids 129-366, which is
responsible for binding with either c-Fos or c-Jun synthesized in
reticulocyte lysate. Next, we located domains within the primary
structure of BAF60a that bind with other SWI·SNF complex subunits.
GST-BAF60a truncation mutants were mixed with cellular lysates prepared
from NIH3T3 cells, subsequently precipitated with GSH-Sepharose, and analyzed by Western blotting with anti-Brm antiserum. As expected, Brm
was precipitated with the full-length BAF60a. Moreover, the same
centrally located domain of BAF60a that binds with c-Fos or c-Jun also
binds with the other subunits of the SWI·SNF complex. We speculate
that these 238 amino acids form a versatile pocket structure and that
any truncation of this region perturbs the three-dimensional structure
of the domain, which leads to a simultaneous loss of binding with the
c-Fos, c-Jun, and SWI·SNF subunits that associate directly with
BAF60a.
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We next asked whether the transactivating activity of c-Fos/c-Jun, the
dimer that binds BAF60a with the highest affinity, is modulated by the
functional SWI·SNF complex in vivo. Assays were performed
with human adenocarcinoma cell line SW13 because it expresses
undetectable levels of Brm and BRG1 (14), which are critical catalytic
(DNA-dependent ATPase) subunits of the SWI·SNF complex. In mammalian
cells, each SWI·SNF complex contains either Brm or BRG1 but not both
(6). Therefore, the introduction of Brm or BRG1 into SW13 would enable
the formation of functional SWI·SNF complex. Transfection of an
expression plasmid encoding either c-Fos, BRG1, or Brm into SW13 cells
did not transactivate a CAT reporter gene coupled to the AP-1 binding
site of the human collagenase gene promoter (Fig.
5A). Transfection of
c-jun alone or c-fos plus c-jun
elevated the CAT activity slightly. Most importantly, transactivation
by c-Fos/c-Jun was strongly potentiated by the addition of Brm.
However, potentiation by BRG1 was marginal. These results suggest that,
in normal cells, transactivation by the c-Fos/c-Jun dimer is strongly
dependent upon the recruitment of the SWI·SNF complex containing Brm
protein.
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We next compared the transactivating potential of Fra-2/JunD, which binds with marginal affinity to BAF60a, with that of c-Fos/c-Jun. Introduction of Fra-2 alone, JunD alone, or Fra2/JunD into SW13 cells did not transactivate the CAT reporter. Moreover, the low CAT activity observed in SW13 cells expressing Fra-2 and JunD was not elevated by the introduction of either Brm or BRG1 (Fig. 5A). These results suggest that a significant portion of the transactivating potential of each Fos/Jun dimer is determined by its binding affinity to BAF60a protein. Without this interaction, the Fos/Jun dimer does not seem to recruit SWI·SNF complex to the AP-1 binding sites. However, the c-Fos/c-Jun dimer retains some transactivating potential even in SW13 cells that lack functional SWI·SNF complex (Fig. 5A). Therefore, there must be some other molecular mechanism that mediates transactivation by the c-Fos/c-Jun dimer.
Considering the possible differences between promoters in transiently
introduced plasmids and native promoters in chromatin, we next examined
the mRNA levels of endogenous collagenase (15) and c-met
(16) genes, which were expected to be inducible through AP-1
binding sites in cells originating from adrenal cortex. Total RNA was
isolated from SW13 cells transfected with the same set of expression
vectors but not with the CAT reporter plasmid. Although each RNA
contained similar amount of GAPDH mRNA as judged by RT-PCR, semiquantitative RT-PCR indicated that the collagenase gene was inducible, dependent on transfection of expression vectors for c-Fos, c-Jun, and Brm (or BRG1) (Fig. 5B). These
results indicate that unlike the results of CAT analysis, BRG1 is
equally efficient for the induction of native collagenase gene
expression. As for the c-met gene, induction is dependent
upon the expression of c-Fos, c-Jun, and BRG1 (Fig. 5B). Brm
has only marginal effects on transactivation of this gene, suggesting
that the Brm or BRG1 subunit has distinct target specificity to
facilitate gene activation. Brm- or BRG1-dependent
transactivation was marginally detected when fra-2 and
junD were transfected instead of c-fos and
c-jun (Fig. 5B).
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DISCUSSION |
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We demonstrated here that SWI·SNF complex subunit BAF60a binds with distinct affinities to different Fos/Jun dimers by interacting with the N-terminal region of c-Jun and the C-terminal region of c-Fos (Fig. 1). BAF60a binds the c-Fos/c-Jun dimer with the highest affinity and the other dimers containing either c-Fos or c-Jun with lower affinity. Fos/Jun dimers that contain neither c-Fos nor c-Jun have no binding activity (Fig. 2). The binding affinity of each Fos/Jun heterodimer to BAF60a is correlated strongly with the respective transactivating activity as determined by transient expression of each heterodimer in F9 cells (Table I). These observations indicate that the SWI·SNF complex is a major determinant of transactivation potential of Fos/Jun dimers. Indeed, in SW13 cells, which lack functional SWI·SNF complex, the transactivating activity of Fos/Jun dimers is kept at basal levels. However, transactivation by c-Fos/c-Jun heterodimer but not by Fra-2/JunD was strongly enhanced by supplying Brm or BRG1 into SW13 to recover the functional SWI·SNF complex (Fig. 5).
Although we did not address the biological function of BAF60a intensively, NIH3T3 cells expressing HA-BAF60a at high levels by retrovirus transduction showed no clear effects on cellular growth. These cells did not acquire anchorage-independent growth at all, indicating that simply a high level expression of BAF60a is not sufficient to induce cellular transformation in NIH3T3 cells (data not shown).
Although several functional models have been presented for the yeast or
mammalian SWI·SNF complex (11), our observations on AP-1 and recent
reports on glucocorticoid receptor (GR) (17-19), c-Myc (20), GCN4 (21,
22), and C/EBP (23) support a model in which transcription
factors recruit the SWI·SNF complex to target genes (24). In these
previous works, INI1 (associates with c-Myc), Brm (associates
with C/EBP
), and Brm or BRG1 (associates with GR) but not BAF60a
were identified as the SWI·SNF complex subunits that bind directly to
transcription factors. These results indicate that the 2-MDa,
multi-subunit SWI·SNF complex has many different interfaces that
interact with various transcription factors and thereby function as a
global transcriptional regulator. Because the transactivating function
of GR in yeast is known to require Swp73p (18), the yeast homologue of
BAF60a, we can speculate that AP-1 (especially c-Fos/c-Jun) and GR
compete for most of the integral subunits of the limited numbers of
SWI·SNF complex present in the cell (25) for their transactivating
activity. Therefore, competition between AP-1 and glucocorticoid
receptor transcription factors for SWI·SNF complex would support in
part the molecular mechanisms for the mutual repression of
transcription frequently observed between these two factors
(26-28).
It is quite interesting that c-Fos and c-Jun are recognized
specifically by BAF60a among Fos family proteins and Jun family proteins, respectively. Both c-fos and c-jun
were isolated originally as cellular counterparts of oncogenes
carried by natural RNA tumor viruses (FBJ-MuSV (12) and ASV17 (29),
respectively), and their products are known to be the strong
transactivators among the family of proteins. Because c-fos
and, to a lesser extent, c-jun are immediate early genes,
the c-Fos/c-Jun dimer would be formed most promptly after extracellular
stimuli and would start to interact with the AP-1 binding site (30,
31). Initially, the chromatin structure would be in an inactive context
for transactivation of AP-1-regulated genes. High affinity binding of
c-Fos/c-Jun with BAF60a efficiently recruits SWI·SNF complex and
thereby induces remodeling of the adjacent chromatin. Subsequently,
basal transcriptional machinery and transactivator proteins such as CBP
and p300 (4, 5) are recruited for the initiation of transcription.
Because the SWI/SNF-remodeled conformation of the nucleosome in some
experimental systems has been shown to persist after detachment of the
SWI·SNF complex (32), it is possible that the promoters within the
modified chromatin remain accessible to regulation by other Fos/Jun
dimers that are induced in later stages, even if these dimers cannot recruit SWI·SNF complex. In summary, the mechanistic link between heterodimeric Fos/Jun transcription factors and chromatin remodeling complexes provided here is essential for understanding the underlying mechanisms that explain the astonishing diversity of AP-1 transcription factor functions in growth, differentiation, development, and tumor formation.
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ACKNOWLEDGEMENTS |
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We thank Dr. H. Kato (Rockefeller University)
for supplying cDNA clones of BRG1 (hSNF2) and Brm (hSNF2
) and
Dr. Y. Miyaji (Toho University) for supplying SW13 cells. We also thank
Dr. M. Brasch (Life Technologies, Inc.) for kind advice on the yeast two-hybrid system (ProQuest Two-Hybrid System), Dr. T. Kameda (Akita
University) for critical reading of the manuscript, and Y. Yoshikawa for preparation of the manuscript.
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FOOTNOTES |
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* This work was supported in part by grants and endowments from Eisai Co., Ltd. and by a grant-in-aid for scientific research on priority areas from the Ministry of Education, Science, and Culture, Japan.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.: 81-3-5449-5730;
Fax: 81-3-5449-5449; E-mail: iba@ims.u-tokyo.ac.jp.
Published, JBC Papers in Press, October 26, 2000, DOI 10.1074/jbc.M009633200
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ABBREVIATIONS |
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The abbreviations used are: PCR, polymerase chain reaction; RT-PCR, reverse transcription-PCR; GSH, gluthathione; GST, glutathione S-transferase; CAT, chloramphenicol acetyltransferase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GR, glucocorticoid receptor.
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REFERENCES |
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1. | Curran, T., and Franza, B. R. (1988) Cell 5, 395-397 |
2. | Nakabeppu, Y., Ryder, K., and Nathans, D. (1988) Cell 55, 907-915[Medline] [Order article via Infotrieve] |
3. | Suzuki, T., Okuno, H., Yoshida, T., Endo, T., Nishina, H., and Iba, H. (1991) Nucleic Acids Res. 19, 5537-5542[Abstract] |
4. | Bannister, A. J., Oehler, T., Wilhelm, D., Angel, P., and Kouzarides, T. (1995) Oncogene 11, 2509-2514[Medline] [Order article via Infotrieve] |
5. | Kamei, Y., Xu, L., Heinzel, T., Torchia, J., Kuroiuwa, R., Gloss, B., Lin, S. C., Heyman, R. A., Rose, D. W., Glass, C. K., and Rosenfeld, M. G. (1996) Cell 85, 403-414[Medline] [Order article via Infotrieve] |
6. | Wang, W., Xue, Y., Zhou, S., Kuo, A., Cairns, B. R., and Crabtree, G. R. (1996) Genes Dev. 10, 2117-2130[Abstract] |
7. | Ui, M., Mizutani, T., Takada, M., Arai, T., Ito, T., Murakami, M., Koike, C., Watanabe, T., Yoshimatsu, K., and Iba, H. (2000) Biochem. Biophys. Res. Commun. 278, 97-105[CrossRef][Medline] [Order article via Infotrieve] |
8. |
Ito, T.,
Kabuyama, Y.,
Okazaki, S.,
Kameda, T.,
Murakami, M.,
and Iba, H.
(1998)
Nucleic Acids Res.
26,
4868-4873 |
9. | Murakami, M., Sonobe, M. H., Ui, M., Kabuyama, Y., Watanabe, H., Wada, T., Handa, H., and Iba, H. (1997) Oncogene 14, 2435-2444[CrossRef][Medline] [Order article via Infotrieve] |
10. | Alani, R., Binetruy, B., Dosaka, H., Rosenberg, R. K., Angle, P., Karin, M., and Birrer, M. J. (1991) Mol. Cell Biol. 11, 6286-6295[Medline] [Order article via Infotrieve] |
11. | Peterson, C. L., and Workman, J. L. (2000) Curr. Opin. Genet. Dev. 10, 187-192[CrossRef][Medline] [Order article via Infotrieve] |
12. | Curran, T., and Teich, N. (1982) J. Virol. 42, 114-122[Medline] [Order article via Infotrieve] |
13. | Curran, T., Miller, A. D., Zokas, L., and Verma, I. M. (1984A) Cell 36, 259-268[Medline] [Order article via Infotrieve] |
14. | Dunaief, J. L., Strober, B. E., Guha, S., Khavari, P. A., Alin, K., Luban, J., Begeman, M., Crabtree, G. R., and Goff, S. P. (1994) Cell 79, 119-130[Medline] [Order article via Infotrieve] |
15. | Gutman, A., and Wasylyk, B. (1990) EMBO J. 9, 2241-2246[Abstract] |
16. | Seol, D. W., Chen, Q., and Zarnegar, R. (2000) Oncogene 19, 1132-1137[CrossRef][Medline] [Order article via Infotrieve] |
17. | Chiba, H., Muramatsu, M., Nomoto, A., and Kato, H. (1994) Nucleic Acids Res. 22, 1815-1820[Abstract] |
18. | Cairns, B. R., Levinson, R. S., Yamamoto, K. R., and Kornberg, R. D. (1996) Genes Dev. 10, 2131-2144[Abstract] |
19. | Fryer, C. J., and Archer, T. K. (1998) Nature 393, 88-91[CrossRef][Medline] [Order article via Infotrieve] |
20. | Cheng, S. W., Davies, K. P., Yung, E., Beltran, R. J., Yu, J., and Kalpana, G. V. (1999) Nat. Genet. 22, 102-105[CrossRef][Medline] [Order article via Infotrieve] |
21. | Natarajan, K., Jackson, B. M., Zhou, H., Winston, F., and Hinnebusch, A. G. (1999) Mol. Cell 4, 657-664[Medline] [Order article via Infotrieve] |
22. | Neely, K. E., Hassan, A. H., Wallberg, A. E., Steger, D. J., Cairns, B. R., Wright, A. P., and Workman, J. L. (1999) Mol. Cell 4, 649-655[Medline] [Order article via Infotrieve] |
23. | Kowenz-Leutz, E., and Leutz, A. (1999) Mol. Cell 4, 735-743[Medline] [Order article via Infotrieve] |
24. |
Yudkovsky, N.,
Logie, C.,
Hahn, S.,
and Peterson, C. L.
(1999)
Genes Dev.
13,
2369-2374 |
25. | Gebuhr, T. C., Bultman, S. J., and Magnuson, T. (2000) Genesis 26, 189-197[CrossRef][Medline] [Order article via Infotrieve] |
26. | Jonat, C., Rahmsdorf, H. J., Park, K. K., Cato, A. C., Gebel, S., Ponta, H., and Herrlich, P. (1990) Cell 62, 1189-1204[Medline] [Order article via Infotrieve] |
27. | Shule, R., Rangarajan, P., Kliewer, S., Ransone, L. J., Bolado, J., Yang, N., Verma, I. M., and Evans, R. M. (1990) Cell 62, 1217-1226[Medline] [Order article via Infotrieve] |
28. | Kameda, T., and Iba, H. (1998) Cancer Res. 58, 867-870[Abstract] |
29. | Maki, Y., Bos, T. J., Davis, C., Starbuck, M., and Vogt, P. K. (1987) Proc. Natl. Acad. Sci. U. S. A. 84, 2848-2852[Abstract] |
30. | Nishina, H., Sato, H., Suzuki, T., Sato, M., and Iba, H. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 3619-3623[Abstract] |
31. | Lallemand, D., Spyrou, G., Yaniv, M., and Pfarr, C. M. (1997) Oncogene 14, 819-830[CrossRef][Medline] [Order article via Infotrieve] |
32. |
Cote, J.,
Peterson, C. L.,
and Workman, J. L.
(1998)
Proc. Natl. Acad. Sci. U. S. A.
95,
4947-4952 |