<i>Drosophila</i> Homologs of the Proto-Oncogene Product PEBP2/CBFβ Regulate the DNA-Binding Properties of Runt

Golling, Gregory; Li, Ling-Hui; Pepling, Melissa E.; Stebbins, Michael; Gergen, J. Peter

doi:10.1128/mcb.16.3.932

Cited by 89 publications

(82 citation statements)

References 47 publications

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“…5,7 Amino acids 1-73 and 102-137 of the 182 amino acid CBF␤ are highly conserved in its two Drosophila homologs, brother and big-brother. 10 Region 1-137 of CBF␤ is sufficient for increasing the DNA affinity of CBF␣ subunits, whereas segment 1-133 does so only weakly. 10,11 The DNA-binding domain of each CBF␣ subunit is located near its N-terminus and is highly homologous to the Drosophila Runt protein.…”

Section: Core Binding Factorsmentioning

confidence: 96%

“…10 Region 1-137 of CBF␤ is sufficient for increasing the DNA affinity of CBF␣ subunits, whereas segment 1-133 does so only weakly. 10,11 The DNA-binding domain of each CBF␣ subunit is located near its N-terminus and is highly homologous to the Drosophila Runt protein. 12 This 128 amino acid 'Runt' domain is responsible for both DNA-binding and heterodimerization with CBF␤.…”

Section: Core Binding Factorsmentioning

confidence: 96%

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1999

Leukemia

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The AML1 and CBF␤ subunits of core binding factor (CBF) are involved in several chromosomal abnormalities frequently associated with acute leukemias. As a result, the CBF␤-SMMHC, AML1-ETO and AML1-MDS1/EVI1 fusion proteins are expressed in subsets of acute myeloid leukemia, and TEL-AML1 is expressed in B-lineage acute lymphocytic leukemia. These CBF oncoproteins likely contribute to leukemogenesis in part by inhibiting endogenous CBF. As a result they are expected to inhibit differentiation and perhaps apoptosis. In addition, the domains unique to each fusion protein may also contribute to leukemogenesis via unique mechanisms.

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Section: Core Binding Factorsmentioning

confidence: 96%

Section: Core Binding Factorsmentioning

confidence: 96%

True

1999

Leukemia

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“…The AML1 protein is a member of the runt family of transcription factors, de®ned by a region of homology to the Drosophila melanogaster segmentation gene runt (termed RHD for runt homology domain) (Daga et al, 1992;Miyoshi et al, 1991), which mediates DNA binding and interacts with Bro and BigBro, which are the Drosophila versions of CBFb (Golling et al, 1996). The consensus DNA sequence recognized by AML1, TGT/cGGT (Meyers et al, 1993), is contained in the promoter and enhancer regions of many hematopoietic-speci®c genes .…”

Section: Introductionmentioning

confidence: 99%

The t(8;21) fusion protein, AML1/ETO, transforms NIH3T3 cells and activates AP-1

et al. 1999

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The 8;21 translocation is the most common cytogenetic abnormality in human acute myelogenous leukemia, joining the AML1 gene on chromosome 21, to the ETO gene on chromosome 8, forming the AML1/ETO fusion gene. The AML1/ETO fusion protein has been shown to function mainly as a transcriptional repressor of AML1 target genes and to block AML1 function in vitro and in vivo. However, AML1/ETO can also activate the BCL-2 promoter and cause enhanced hematopoietic progenitor self-renewal in vitro, suggesting gain-of-functions unique to the fusion protein. We used NIH3T3 cells to determine the transforming capacity of AML1/ETO, and to further characterize its mechanism of action. Expression of AML1/ETO in NIH3T3 cells caused cell-type speci®c cell death, and cellular transformation, characterized by phenotypic changes, anchorage-independent growth, and tumor formation in nude mice. In contrast, neither expression of AML1A, AML1B or ETO altered the normal growth pattern of the cells. To investigate the mechanism of transformation by AML1/ETO, we analysed the levels of activated, phosphorylated c-Jun (ser63) and other constituents of the AP-1 complex, in the presence of various AML1/ ETO related proteins. Expression of AML1/ETO increased the level of c-Jun-P (ser63), and activated AP-1 dependent transcription, which was inhibited by expression of a dominant-negative c-Jun protein. Mutational analysis revealed that the runt homology domain (RHD) and a C-terminal transcriptional repression domain in AML1/ETO are required for transformation, activation of c-Jun and increased AP-1 activity. These results establish the transforming potential of the t(8;21) fusion protein and link this gain-of-function property to modulation of AP-1 activity.

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“…2g). The Runt/Bgb complex binds efficiently to this recognition sequence in vitro, whereas Runt in the absence of Bgb has a lower affinity for its target sequence 21 . Activation of a reporter containing a representative CBFα/β binding site is dependent on the presence of both members of the heterodimer (Supplementary Fig.…”

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confidence: 99%

A bacterial one-hybrid system for determining the DNA-binding specificity of transcription factors

2005

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The DNA-binding specificities of transcription factors can be used to computationally predict cisregulatory modules (CRMs) that regulate gene expression 1 . However, the absence of specificity data for the majority of transcription factors limits the wide-spread implementation of this approach. We have developed a bacterial one-hybrid system that provides a simple and rapid method to determine the DNA-binding specificity of a transcription factor. Using this technology, we successfully determined the DNA-binding specificity of seven previously characterized transcription factors and one novel transcription factor, the Drosophila factor Odd-skipped. Regulatory targets of Odd-skipped were successfully predicted using this information, demonstrating that the data produced by the bacterial one-hybrid system is relevant to in vivo function.Several methods exist for determining the DNA-binding specificity of a transcription factor. SELEX is the most commonly employed method to define DNA-binding specificity 2 . More recently, microarrays of short oligonucleotides 3 or intergenic sequences 4 have been used to characterize transcription factors. Genome-wide chromatin immunoprecipitation (ChIPchip) 5 and DNA immunoprecipitation with microarray detection (DIP-chip) 6 have also been used in conjunction with computational analysis to identify statistically overrepresented sequence motifs to extract DNA-binding specificity from the genomic segments that are bound by a transcription factor. These methods, while powerful, have drawbacks: the in vitro technologies require the purification of the transcription factor in its active form; SELEX requires multiple rounds of selection to complete; and microarray-based techniques require the facilities and expertise to analyze the arrays and resulting data.We have developed a bacterial one-hybrid (B1H) system that provides a simple method for defining the DNA-binding specificity of a transcription factor. The selection procedure is rapid, because only a single round of selection is required to generate a set transcription factor binding sites, and it is readily accessible, because only basic molecular biology expertise is required to employ the technology. Conceptually, this system is similar to yeast one-hybrid systems that can determine the DNA-binding specificity of a transcription factor 7 and detect protein-DNA interactions 8 . However, a bacterial selection system provides advantages over the

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Drosophila Homologs of the Proto-Oncogene Product PEBP2/CBFβ Regulate the DNA-Binding Properties of Runt

Cited by 89 publications

References 47 publications

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The t(8;21) fusion protein, AML1/ETO, transforms NIH3T3 cells and activates AP-1

A bacterial one-hybrid system for determining the DNA-binding specificity of transcription factors

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