A Dominant Negative to Activation Protein-1 (AP1) That Abolishes DNA Binding and Inhibits Oncogenesis

Olive, Michelle; Krylov, Dmitry; Echlin, Deborah R.; Gardner, Kevin; Taparowsky, Elizabeth J.; Vinson, Charles

doi:10.1074/jbc.272.30.18586

Cited by 266 publications

(314 citation statements)

References 45 publications

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“…Cotransfection of KCREB with the CRE reporter caused a threefold increase in CRE transcriptional activity (Figure 7a). Co-transfection of the CRE reporter with A-CREB caused a 7 ± 10-fold increase in transcriptional activity whereas A-ZIP DN constructs speci®c for the Fos (A-FOS) and C/EBP (A-C/EBP) (Olive et al, 1997;Ahn et al, 1998) families of transcription factors did not alter CRE reporter activity (Figure 7b). Constructs encoding either the CREB leucine zipper but not the acidic extension (OH-CREB), or the wildtype CREB C-terminus, leucine zipper and basic region, but not the transactivation domain (B-CREB) failed to alter the activity of the CRE reporter ( Figure 7b) indicating that the acidic extension is required for the observed e ect of the A-CREB mutant.…”

Section: Depression Of Cre Transcriptional Activity In Keratinocytesmentioning

confidence: 99%

“…In a second approach to examine the potential involvement of CREB family proteins in the AP-1 DNA binding complex, we have included dominantnegative (DN) mutant proteins (described in Olive et al, 1997;Ahn et al, 1998) which inhibit the binding of Fos family (A-Fos), CREB family (A-CREB) and VBP (A-VBP) proteins to DNA in the binding reactions. The addition of the A-FOS or A-CREB mutants to the reaction decreased the binding of the extracts to the 32 P-labeled AP-1 sequence to a similar extent ( Figure 1b).…”

Section: Activities Of Ap-1 and Cre Dna Binding Proteins In Keratinocmentioning

confidence: 99%

“…The A-CREB construct encodes a protein containing the CREB leucine zipper and Cterminus. In this case, the CREB DNA binding region has been replaced by an amphipathic acidic extension designed to stabilize the formation of the heterodimer while preventing interactions with DNA (as described in Krylov et al, 1995;Olive et al, 1997). Cotransfection of KCREB with the CRE reporter caused a threefold increase in CRE transcriptional activity (Figure 7a).…”

Section: Depression Of Cre Transcriptional Activity In Keratinocytesmentioning

confidence: 99%

“…The wild-type and mutated forms of the TGFb 3 promoter (Lafyatis et al, 1990) were obtained from S Kim (National Cancer Institute, Bethesda, MD, USA). CMV-500 and the A-ZIP vectors CMV-A-CREB, CMV-A-FOS, CMV-A-C/EBP, CMV-B-CREB and CMV-OH-CREB vectors have been described (Olive et al, 1997;Krylov et al, 1995;Ahn et al, 1998). The RSV CREMa and RSVKCREB expression vectors were obtained from R Goodman (Oregon Health Sciences University, Portland, OR, USA), the CMV-Fra-2 construct was obtained from Donna Cohen (The Australian University, Canberra, Australia).…”

Section: Plasmids Proteins and Antibodiesmentioning

confidence: 99%

“…This study examines the expression patterns and DNA binding activities of CREB, CREMa and ATF-1 in basal and di erentiating keratinocytes. Using dominant-negative (DN) mutants (A-ZIP) mutants Ahn et al, 1998, Olive et al, 1997 that dimerize with the CREB and Fos families of transcription factors and inhibit their ability to bind to DNA, we have established that both Fos and CREB proteins bind to the AP-1 target sequence and suppress AP-1 transcriptional activity in primary cultures of di erential keratinocytes.…”

Section: Introductionmentioning

confidence: 99%

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CRE DNA binding proteins bind to the AP-1 target sequence and suppress AP-1 transcriptional activity in mouse keratinocytes

et al. 1999

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Previously, we have shown that nuclear extracts from cultured mouse keratinocytes induced to di erentiate by increasing the levels of extra-cellular calcium contain Fra-1, Fra-2, Jun B, Jun D and c-Jun proteins that bind to the AP-1 DNA binding sequence. Despite this DNA binding activity, AP-1 reporter activity was suppressed in these cells. Here, we have detected the CREB family proteins CREB and CREMa as additional participants in the AP-1 DNA binding complex in di erentiating keratinocytes. AP-1 and CRE DNA binding activity correlated with the induction of CREB, CREMa and ATF-1 and CREB phosphorylation at ser 133 (ser 133 phospho-CREB) in the transition from basal to di erentiating keratinocytes, but the activity of a CRE reporter remained unchanged. In contrast, the CRE reporter was activated in the presence of the dominantnegative (DN) CREB mutants, KCREB and A-CREB, proteins that dimerize with CREB family members and block their ability to bind to DNA. The increase in CRE reporter activity in the presence of these mutants suggests that CRE-mediated transcriptional activity is suppressed in keratinocytes through protein-protein interactions involving a factor that dimerizes with the CREB leucine zipper. In experiments where the A-CREB mutant was co-transfected with an AP-1 reporter construct, transcriptional activity was also increased indicating that a CREB family member binds AP-1 sites and represses AP-1 transcriptional activity as well. Exogenous expression of the transcriptional repressor CREMa down-regulated both CRE and AP-1 reporters in keratinocytes suggesting that this factor may contribute to the suppression of AP-1 transcriptional activity observed in di erentiating keratinocytes.

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Section: Depression Of Cre Transcriptional Activity In Keratinocytesmentioning

confidence: 99%

Section: Activities Of Ap-1 and Cre Dna Binding Proteins In Keratinocmentioning

confidence: 99%

Section: Depression Of Cre Transcriptional Activity In Keratinocytesmentioning

confidence: 99%

Section: Plasmids Proteins and Antibodiesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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CRE DNA binding proteins bind to the AP-1 target sequence and suppress AP-1 transcriptional activity in mouse keratinocytes

et al. 1999

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Endocrine response and resistance in breast cancer: a role for the transcription factor Fos

Gee¹,

Willsher²,

Kenny³

et al. 1999

Int. J. Cancer

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We have previously demonstrated that elevated Fos expression may be important in de novo endocrine resistance in breast cancer. However, changes in Fos expression during endocrine response and subsequently on acquisition of resistance are unknown. This study immunocytochemically monitors Fos protein within sequential biopsies from primary human breast cancer patients obtained pre‐treatment (T1), during tamoxifen therapy (T2, T3) and on disease progression (T5), examining in parallel proliferation [i.e., MIB1 (Ki67) immunostaining, mitotic activity], cellularity and endocrine response. Significantly diminished Fos, proliferation and cellularity were observed after 6 weeks of therapy in patients exhibiting a better quality and/or duration of response, while modest Fos increases and a maintained proliferation and cellularity were seen in poorer responders. Decreases in Fos, proliferation and cellularity at 6 months similarly hallmarked better responders. We confirmed a significant association between de novo resistance and elevated Fos and proliferation. Additionally, however, these parameters increased at the time of disease relapse over pre‐treatment and “on therapy” values. Our data indicate that tamoxifen response involves a reduction in both tumor cell proliferation and cell survival, potentially entailing diminished Fos protein expression in better‐responding patients. Our data are also supportive of elevated Fos expression being involved in the departure from endocrine control inherent in both primary and acquired resistance. Int. J. Cancer (Pred. Oncol.) 84:54–61, 1999. © 1999 Wiley‐Liss, Inc.

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Uncoiling c-Jun coiled coils: Inhibitory effects of truncated Fos peptides on Jun dimerization and DNA binding in vitro

et al. 1998

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c-Jun is an oncoprotein that comprises a portion of the AP-1 transcription factor and belongs to the basic-leucine zipper (bZIP) DNA binding protein family. Using peptides derived from the leucine zipper region of Fos, we have developed agents that inhibit Jun's DNA binding in the low micromolar range. Fos peptides that were effective inhibitors in the DNA binding assay were also found to inhibit cellular Jun binding to an AP-1 site in a luciferase reporter plasmid in MCF-7 cells. Size exclusion studies confirmed that peptides that inhibit the DNA binding of Jun also inhibit its dimerization. These peptides were found to have a cytotoxic effect on the MCF-7 cell line when delivered with the transfecting agent Tfx-50, possibly due to their role as transcription factor regulators.

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A Dominant Negative to Activation Protein-1 (AP1) That Abolishes DNA Binding and Inhibits Oncogenesis

Cited by 266 publications

References 45 publications

CRE DNA binding proteins bind to the AP-1 target sequence and suppress AP-1 transcriptional activity in mouse keratinocytes

CRE DNA binding proteins bind to the AP-1 target sequence and suppress AP-1 transcriptional activity in mouse keratinocytes

Endocrine response and resistance in breast cancer: a role for the transcription factor Fos

Uncoiling c-Jun coiled coils: Inhibitory effects of truncated Fos peptides on Jun dimerization and DNA binding in vitro

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