Modulation of Transcription Factor Ets-1 DNA Binding: DNA-Induced Unfolding of an α Helix

Petersen, Jeannine M.; Skalicky, Jack J.; Donaldson, Logan W.; McIntosh, Lawrence P.; Alber, Tom; Graves, Barbara J.

doi:10.1126/science.7569926

Cited by 175 publications

(217 citation statements)

References 41 publications

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“…Based on NMR criteria, the helical content of Ets-1 AN280 is approximately 35%, which is similar to that observed previously for Ets-lAN331 (Donaldson et al, 1994). This is consistent with CD measurements showing that Ets-lAN280 and Ets-lAN331 have similar mean residue molar ellipticities [a] at 222 nm (Petersen et al, 1995).…”

Section: Secondary Structure Of Ets-im280: a Whth Motif Flanked By Hesupporting

confidence: 89%

“…Deletion of either region results in as much as a IO-20-fold increase in affinity for DNA (Hagman & Grosschedl, 1992;Lim et al, 1992;Nye et al, 1992;Wasylyk et al, 1992;Fisher et al, 1994;Petersen et al, 1995;Jonsen et al, 1996). Several natural variants of Ets-1, differing in either the amino or carboxy sequences flanking the ETS domain, also display enhanced DNA binding.…”

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confidence: 99%

“…Ets-IAN280 contains residues 280-440 of Ets-I, including the ETS domain and both flanking inhibitory regions. Quantitative binding studies demonstrate that Ets-lAN280 and full-length Ets-1 are inhibited to a similar degree and make identical contacts to DNA (Petersen et al, 1995; J.M. Petersen, Q.-P. Xu, B. J. Graves, unpubl.…”

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confidence: 99%

“…In addition to the three a-helices and four @-strands in the wHTH ETS domain, Ets-lAN280 is comprised of two a-helices and an unstructured region in the amino-terminal inhibitory sequence and one a-helix in the carboxy-terminal sequence. A comparison of the backbone chemical shifts of Ets-IAN280 were made with those of a high-affinity DNAbinding fragment, Ets-IAN33 1, that lacks the amino-terminal inhibitory residues (Donaldson et al, 1994;Petersen et al, 1995). The spectral differences demonstrate that the amino-and carboxy-terminal inhibitory sequences are structurally coupled and that they interact with the ETS domain.…”

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confidence: 99%

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Structural coupling of the inhibitory regions flanking the ETS domain of murine Ets‐1

et al. 1996

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Several members of the ets gene family of transcription factors show negative regulation of DNA binding by intramolecular interactions. A structural mechanism for this auto-inhibition is investigated using a 161-residue N-terminal deletion mutant of murine Ets-1, Ets-1AN280. This protein shows a similar reduced affinity for DNA as native Ets-1 because it contains the ETS domain in context of the flanking amino-and carboxy-terminal regions that together mediate repression of DNA binding. The secondary structure of Ets-IAN280 was determined using NMR chemical shift, NOE, J coupling, and amide hydrogen exchange information. In addition to the winged helix-turn-helix ETS domain, Ets-1AN280 contains two a-helices in the amino-terminal inhibitory region and one a-helix in the carboxy-terminal inhibitory region. Chemical shift comparisons were made between this protein and an activated form of Ets-1 lacking the amino-terminal inhibitory region. The spectral differences demonstrate that the amino-and carboxy-terminal inhibitory sequences are structurally coupled to one another, thus explaining the observation that both regions are required for the repression of DNA binding. Furthermore, these data show that the inhibitory sequences also interact directly with the first helix of the intervening ETS domain, thereby providing a pathway for the repression of DNA binding. These results lead to a model of an inhibitory module in Ets-1 composed of both the amino-and carboxy-terminal regions interfaced with the ETS domain. This establishes the structural framework for understanding the intramolecular inhibition of Ets-1 DNA binding.Keywords: allosteric; DNA binding; ETS domain; Ets-1 and Ets-2; intramolecular inhibition; NMR; protein structure; winged helix-turn-helix DNA binding, activation, and oligomerization associated with a particular transcription factor may be governed by its subcellular localization, posttranslational modifications, ligand binding, and protein-protein interactions.Recently, data have Abbreviations: ets, E26 transformation specific; Ets-1AC428, the carboxy-terminal deletion fragment of murine Ets-1 consisting of residues 1-428; Ets-1AN280, the amino-terminal deletion fragment of murine Ets-1 consisting of residues 280-440; Ets-1AN331, the aminoterminal deletion fragment of murine Ets-1 consisting of residues 33 1-440; FID, free induction decay; HSQC, heteronuclear single quantum correlation; NOESY, nuclear Overhauser effect spectroscopy; TOCSY, total correlation spectroscopy; IPTG, isopropyl-8-D-thiogalactopyranoside; DTT, dithiothreitol; pH*, the observed pH meter reading without correction for isotope effects; wHTH, winged helix-turn-helix; HSF, heat shock transcription factors; DSS, 2,2-dimethyl-2-silapentane-5-sulfonate, sodium salt.

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Section: Secondary Structure Of Ets-im280: a Whth Motif Flanked By Hesupporting

confidence: 89%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Structural coupling of the inhibitory regions flanking the ETS domain of murine Ets‐1

et al. 1996

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“…The following possibilities for such interplay should be considered. First, the functions of ETS proteins are often associated with the binding of cooperative regulatory proteins which could stabilize the interactions between the ETS and DNA (Petersen et al, 1995) and are necessary for speci®c gene transactivation in particular cell types (reviewed in: Papas et al, 1997). Second, the oncogenic conversion of Fli-1, Erg and other Ets related genes via gene fusion has been reported in EWS cells (Delattre et al, 1992;May et al, 1993;Sorensen et al, 1994;Jeon et al, 1995;Peter et al, 1997).…”

Section: Inhibition Of Ets1 Expression Enhances Growth Retardation Ofmentioning

confidence: 99%

Regulation of the human poly(ADP-ribose) polymerase promoter by the ETS transcription factor

et al. 1999

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Ewing's sarcoma (EWS) cells accumulate elevated steady-state levels of poly (ADP-ribose) polymerase (PARP) mRNA and protein. To understand the molecular mechanisms underlying PARP upregulation, we cloned and analysed the 5'-¯anking region of the PARP gene from EWS cells. Nucleotide sequence analysis demonstrated no variations in the PARP promoter region in EWS cells. The PARP promoter encompasses multiple binding motifs for the ETS transcription factor. We have also observed that there is a coordinated up-regulation of the expression of both PARP and ETS1, relative to cells of other human tumor types expressing lower levels of PARP. Transient coexpression of ETS1 in EWS cells resulted in a strong enhancement of PARP-promoter activity. The participation of ETS in the regulation of PARP gene expression was further demonstrated in EWS cells stably transfected with Ets1 antisense cDNA constructs. Antisensemediated down-regulation of endogenous ETS1 resulted in the inhibition of PARP expression in EWS cells, and sensitized these cells to ionizing radiation. These data provide support for ETS regulation of PARP expression levels, and implicate ETS transcription factors in the radiation response of EWS cells.

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Upregulation of human heme oxygenase gene expression by Ets-family proteins

1999

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Overexpression of human heme oxygenase-1 has been shown to have the potential to promote EC proliferation and angiogenesis. Since Ets-family proteins have been shown to play an important role in angiogenesis, we investigated the presence of ETS binding sites (EBS), GGAA/T, and ETS protein contributing to human HO-1 gene expression. Several chloramphenicol acetyltransferase constructs were examined in order to analyze the effect of ETS family proteins on the transduction of HO-1 in Xenopus oocytes and in microvessel endothelial cells. Heme oxygenase promoter activity was up-regulated by FLI-1ERGETS-1 protein(s). Chloramphenicol acetyltransferase (CAT) assays demonstrated that the promoter region (-1500 to +19) contains positive and negative control elements and that all three members of the ETS protein family were responsible for the up-regulation of HHO-1. Electrophoretic mobility shift assays (EMSA), performed with nuclear extracts from endothelial cells overexpressing HHO-1 gene, and specific HHO-1 oligonucleotides probes containing putative EBS resulted in a specific and marked bandshift. Synergistic binding was observed in EMSA between AP-1 on the one hand, FLI-1, ERG, and ETS-1 protein on the other. Moreover, 5'-deletion analysis demonstrated the existence of a negative control element of HHO-1 expression located between positions -1500 and -120 on the HHO-1 promoter. The presence of regulatory sequences for transcription factors such as ETS-1, FLI-1, or ERG, whose activity is associated with cell proliferation, endothelial cell differentiation, and matrix metalloproteinase transduction, may be an indication of the important role that HO-1 may play in coronary collateral circulation, tumor growth, angiogenesis, and hemoglobin-induced endothelial cell injuries.

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Modulation of Transcription Factor Ets-1 DNA Binding: DNA-Induced Unfolding of an α Helix

Cited by 175 publications

References 41 publications

Structural coupling of the inhibitory regions flanking the ETS domain of murine Ets‐1

Structural coupling of the inhibitory regions flanking the ETS domain of murine Ets‐1

Regulation of the human poly(ADP-ribose) polymerase promoter by the ETS transcription factor

Upregulation of human heme oxygenase gene expression by Ets-family proteins

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