Isolation of transcription factors that discriminate between different promoters recognized by RNA polymerase II

Dynan, William S.; Tjian, Robert

doi:10.1016/0092-8674(83)90053-3

Cited by 745 publications

(375 citation statements)

References 32 publications

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“…Competition might be for RNA polymerase II directly or for one of several potentially rate-limiting transcription factors already identified in HeLa cells (Dynan and Tjian, 1983). The relevance of these observations to the pattern of expression of the -y and ,3 genes in fetal erythroid cells is conjectural but several facts are of interest.…”

Section: Discussionmentioning

confidence: 95%

Promoter sequences required for function of the human gamma globin gene in erythroid cells.

Anagnou¹,

Karlsson²,

Moulton³

et al. 1986

The EMBO Journal

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The human 'y-and 3-globin genes are expressed during the fetal and adult developmental periods, respectively. Differences in the sequences of their promoters may be relevant to their developmental regulation. The y globin gene promoter was found to be stronger than that of the , gene. In HeLa cells co-transfected with plasmids containing the two genes, transcripts arising from the -y promoter accumulated to a level 3-fold higher than those initiated from the f-promoter. We have recently shown that deletion of the most distal of the conserved elements of the promoter (CACCC) reduces function to 25% of the wild-type, whereas the removal of the proximal of the two duplicated (CCAAT) elements increases promoter function 2-to 4-fold in HeLa cells during transient gene expression. Both the wild-type promoter and the truncation and linker-scanning mutants from which one of the two duplicated 'CCAAT' elements had been removed, exhibited regulated expression when stably integrated into chromosomes of mouse erythroleukemia (MEL) cells. Thus, duplication of the 'CCAAT' element, the feature by which the 'y promoter differs most strikingly from the (3, is not essential for function in erythroid cells.

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Section: Discussionmentioning

confidence: 95%

Promoter sequences required for function of the human gamma globin gene in erythroid cells.

Anagnou¹,

Karlsson²,

Moulton³

et al. 1986

The EMBO Journal

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“…Sp1 specifically interacts with DNA regions containing defined GC boxes to activate transcription (8). Distinct regions of this protein have been shown to direct DNA binding and transcriptional activation.…”

Section: Resultsmentioning

confidence: 99%

Myogenic Basic Helix-Loop-Helix Proteins and Sp1 Interact as Components of a Multiprotein Transcriptional Complex Required for Activity of the Human Cardiac α-Actin Promoter

Biesiada

Hamamori

Kedes

et al. 1999

Molecular and Cellular Biology

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Activation of the human cardiac ␣-actin (HCA) promoter in skeletal muscle cells requires the integrity of DNA binding sites for the serum response factor (SRF), Sp1, and the myogenic basic helix-loop-helix (bHLH) family. In this study we report that activation of the HCA correlates with formation of a muscle-specific multiprotein complex on the promoter. We provide evidence that proteins eluted from the multiprotein complex specifically react with antibodies directed against myogenin, Sp1, and SRF and that the complex can be assembled in vitro by using the HCA promoter and purified MyoD, E12, SRF, and Sp1. In vitro and in vivo assays revealed a direct association of Sp1 and myogenin-MyoD mediated by the DNA-binding domain of Sp1 and the HLH motif of myogenin. The results obtained in this study indicate that protein-protein interactions and the cooperative DNA binding of transcriptional activators are critical steps in the formation of a transcriptionally productive multiprotein complex on the HCA promoter and suggest that the same mechanisms might be utilized to regulate the transcription of muscle-specific and other genes.Cooperative interactions of transcriptional activators are pivotal in ensuring the proper execution of the myogenic program. For instance, the cooperative binding to two adjacent E boxes on the muscle creatine kinase enhancer by MyoD is required for transcriptional activation (38). The transcriptional activators that bind muscle regulatory regions often establish direct contacts. In fact, protein-protein interactions govern functional cooperativity of myogenic basic helix-loop-helix (bHLH) with E proteins (17) and the myocyte enhancer factor 2 (MEF2) in directing muscle transcription (24). In addition to the E box, numerous muscle-specific regulatory regions contain binding sites for the MEF2 proteins, the serum response factor (SRF), and Sp1, suggesting that the combinatorial binding of these factors to muscle regulatory regions has been selected and is particularly favored for regulation of musclespecific transcription (25, 39). Whereas both MEF2 and SRF have been shown to interact with the myogenic bHLH (24, 11), the question of whether Sp1 can also directly associate with myogenic bHLH has not been addressed. Furthermore, it remains to be determined whether muscle and ubiquitous transcription factors found on muscle-specific regulatory regions are associated as multiprotein complexes.A region of the human cardiac ␣-actin (HCA) promoter spanning from nucleotides Ϫ110 to ϩ68 is a composite response element that directs striated muscle-specific transcription (20). Electrophoretic mobility shift assay (EMSA) and footprinting experiments revealed sites for the binding of three nuclear proteins in myogenic cells, including a CArG box for the SRF (21, 5), a GC box for Sp1 (12), and an E box for one of the myogenic bHLH proteins (33). Mutations in any of these three DNA elements result in promoter inactivation (33). Furthermore, reconstitution experiments conducted in Drosophila melanogaster-deriv...

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“…There are several lines of evidence suggesting that an increased amount of Spl protein would have profound effects on cellular transcriptional activity and aid in viral transcription. In vitro assays using the SV40 promoter (Dynan and Tjian 1983a) and a monkey promoter {Dynan et al 1985) have shown that transcription rates increase with higher amounts of Sp 1. A similar dependence is seen in assays in vivo (Courey and Tjian 1988).…”

Section: Spl Activation By Sv40mentioning

confidence: 99%

SV40 stimulates expression of the transacting factor Sp1 at the mRNA level.

Saffer¹,

Jackson²,

Thurston³

1990

Genes Dev.

123

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Isolation of transcription factors that discriminate between different promoters recognized by RNA polymerase II

Cited by 745 publications

References 32 publications

Promoter sequences required for function of the human gamma globin gene in erythroid cells.

Promoter sequences required for function of the human gamma globin gene in erythroid cells.

Myogenic Basic Helix-Loop-Helix Proteins and Sp1 Interact as Components of a Multiprotein Transcriptional Complex Required for Activity of the Human Cardiac α-Actin Promoter

SV40 stimulates expression of the transacting factor Sp1 at the mRNA level.

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