The Structure of GABPα/β: An ETS Domain- Ankyrin Repeat Heterodimer Bound to DNA

Batchelor, Adrian H.; Piper, Derek E.; Brousse, Fabienne Charles de la; McKnight, Steven L.; Wolberger, Cynthia

doi:10.1126/science.279.5353.1037

Cited by 278 publications

(256 citation statements)

References 26 publications

(13 reference statements)

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“…Therefore, GABPb or GABPg should bind GABPa in the cytoplasm to assist GABPa transport into the nucleus. [18][19][20] GABP is now also recognized as a key transcriptional regulator of dynamically regulated, lineage-restricted genes, especially in myeloid cells and at the neuromuscular junction, and acts as an integrator of the cellular signaling pathway by regulating key hormones and transmembrane receptors. 17 Recently, we found that the heteromeric complex of GABP (GABPa/b) acts as a transcriptional repressor and thereby inhibits VEGF expression in SH-SY5Y neuroblastoma and green monkey kidney epithelial (CV-1) cells.…”

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confidence: 99%

Subconjunctival gene delivery of the transcription factor GA-binding protein delays corneal neovascularization in a mouse model

et al. 2009

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Corneal neovascularization can reduce visual acuity. GAbinding protein (GABP) is a transcription factor that regulates the expression of target genes including vascular endothelial growth factor (VEGF) and roundabout4 (Robo4), which participate in pathologic angiogenesis. We assessed whether intraocular injection of the GABP gene affects the growth of new corneal blood vessels in a mouse ocular neovascularization model. Transfection of human GABPa and GABPb gene (GABPa/b) into human conjunctival epithelial cells resulted in decreased VEGF and Robo4 expression. Three groups of mice underwent chemical and mechanical denudation of the corneal epithelium. Subsequently, two groups were administered subconjunctival injection of lipoplexes carrying plasmid DNA encoding for human GABPa/b or an empty plasmid DNA at 1-week intervals. The third group served as an experimental control. In vivo delivery of human GABPa/b into mouse neovascularized cornea reduced VEGF and Robo4 gene expression. Biomicroscopic examination showed that, at 1 week after one or two injections, GABPa/b-treated eyes had significantly less neovascularized corneal area than did eyes treated with the empty vector. Histologic examination showed significantly less vascularized area and fewer blood vessels in the GABP-treated group at 1 week after injections. However, these angiosuppressive effects were weakened at 2 weeks after injections. Our results indicate that subconjunctival GABP gene delivery delays corneal neovascularization for up to 2 weeks in a mouse model of deliberate corneal injury.

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Section: Introductionmentioning

confidence: 99%

Subconjunctival gene delivery of the transcription factor GA-binding protein delays corneal neovascularization in a mouse model

et al. 2009

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“…Upon the basis of NMR analyses of the secondary structures of tumor suppressor p16, we first reported that ankyrin-repeats consist of helix-turn-helix (H-T-H) motifs (13). Subsequently, the tertiary structures of several proteins containing ankyrin repeats have been reported: the crystal structures of 53BP2 (14), p18 (15), and transcriptional regulator GABPR/ bound to DNA (16) and the solution structures of p19 (17), p16 (18), and myotrophin (19). In all of these structures, ankyrin repeats exist as H-T-H motifs linked by loops.…”

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confidence: 99%

“…(b) In the crystal structure of the transcriptional regulator GABPR/ -DNA complex, the -subunit consists of 5 ankyrin repeats linked by 4 loops. The interface between R-and -subunits involves mainly the tips of the loops of the -subunit (16). The tip of one of the loops in 53BP2 is also involved in the binding to p53 (14).…”

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confidence: 99%

“…(a) It has been suggested that the loops (18) or the tips of the loops (16) are the possible sites for protein-protein interactions, including the interaction between p16 and CDK4, which is of enormous significance in cancer research. (b) In the crystal structure of the transcriptional regulator GABPR/ -DNA complex, the -subunit consists of 5 ankyrin repeats linked by 4 loops.…”

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confidence: 99%

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Tumor Suppressor INK4: Determination of the Solution Structure of p18^INK4C and Demonstration of the Functional Significance of Loops in p18^INK4C and p16^INK4A ^,

Byeon

Ericson

et al. 1999

Biochemistry

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Since the structures of several ankyrin-repeat proteins including the INK4 (inhibitor of cyclindependent kinase 4) family have been reported recently, the detailed structures and the functional roles of the loops have drawn considerable interest. This paper addresses the potential importance of the loops of ankyrin-repeat proteins in three aspects. First, the solution structure of p18 INK4C was determined by NMR, and the loop structures were analyzed in detail. The loops adapt nascent antiparallel -sheet structures, but the positions are slightly different from those in the crystal structure. A detailed comparison between the solution structures of p16 and p18 has also been presented. The determination of the p18 solution structure made such detailed comparisons possible for the first time. Second, the { 1 H, 15 N}HSQC NMR experiment was used to probe the interactions between p18 INK4C and other proteins. The results suggest that p18 INK4C interacts very weakly with dna K and glutathione S-transferase via the loops. The third aspect employed site-specific mutagenesis and functional assays. Three mutants of p18 and 11 mutants of p16 were constructed to test functional importance of loops and helices. The results suggest that loop 2 is likely to be part of the recognition surface of p18 INK4C or p16 INK4A for CDK4, and they provide quantitative functional contributions of specific residues. Overall, our results enhance understanding of the structural and functional roles of the loops in INK4 tumor suppressors in particular and in ankyrinrepeat proteins in general.The structure and function of the INK4 family of tumor suppressors are active subjects of current research in many laboratories. There are four known members in this family: p15 INK4B

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“…The DNA-binding component is the ␣ subunit (GABP␣) containing the conserved Ets domain, a winged helix-turn-helix protein motif (6). The ␤ subunit (GABP␤) is bound to GABP␣ via repeats of the Notch-ankyrin type (19).…”

mentioning

confidence: 99%

Synergistic Action of GA-Binding Protein and Glucocorticoid Receptor in Transcription from the Mouse Mammary Tumor Virus Promoter

Aurrekoetxea-Hernández

Buetti

2000

J Virol

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B lymphocytes are among the first cells to be infected by mouse mammary tumor virus (MMTV), and they play a crucial role in its life cycle. To study transcriptional regulation of MMTV in B cells, we have analyzed two areas of the long terminal repeat (LTR) next to the glucocorticoid receptor binding site, fp1 (at position ؊139 to ؊146 from the cap site) and fp2 (at ؊157 to ؊164). Both showed B-cell-specific protection in DNase I in vitro footprinting assays and contain binding sites for Ets transcription factors, a large family of proteins involved in cell proliferation and differentiation and oncogenic transformation. In gel retardation assays, fp1 and fp2 bound the heterodimeric Ets factor GA-binding protein (GABP) present in B-cell nuclear extracts, which was identified by various criteria: formation of dimers and tetramers, sensitivity to pro-oxidant conditions, inhibition of binding by specific antisera, and comigration of complexes with those formed by recombinant GABP. Mutations which prevented complex formation in vitro abolished glucocorticoid-stimulated transcription from an MMTV LTR linked to a reporter gene in transiently transfected B-cell lines, whereas they did not affect the basal level. Exogenously expressed GABP resulted in an increased level of hormone response of the LTR reporter plasmid and produced a synergistic effect with the coexpressed glucocorticoid receptor, indicating cooperation between the two. This is the first example of GABP cooperation with a steroid receptor, providing the opportunity for studying the integration of their intracellular signaling pathways.Mouse mammary tumor virus (MMTV) is a type B retrovirus which causes carcinomas of the mammary gland in females of susceptible mouse strains (8, 13). The carcinogenic transformation results from transcriptional activation of a particular class of cellular oncogenes (int genes; reviewed in reference 59). The mammary gland specificity of the oncogenic property of MMTV depends on the high viral replication rate and consequent high reinfection rate in the mammary epithelial cells, which are stimulated by pregnancy-related hormones. This hormonal stimulation has made the MMTV promoter the beststudied model for investigating the regulation of gene expression by steroid hormones (for a review, see references 7 and 51). Glucocorticoids, progesterone, and androgens (but not estrogens) strongly stimulate the rate of MMTV transcription through the binding of hormone-receptor complexes in the hormone regulatory element (HRE) (see Fig. 1) region of the proviral DNA, located upstream of the promoter in the long terminal repeat (LTR). Binding sites for additional transcription factors have been characterized, in particular for CTF (also called nuclear factor 1 [NF-1]) and Oct-1 in the HRE, and for mammary gland-specific factors further upstream in the LTR (reviewed in reference 31).During primary infection, MMTV is transmitted in the milk from the mother to the newborn and is taken up in the intestine, where it infects local lymphocytes (rev...

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The Structure of GABPα/β: An ETS Domain- Ankyrin Repeat Heterodimer Bound to DNA

Cited by 278 publications

References 26 publications

Subconjunctival gene delivery of the transcription factor GA-binding protein delays corneal neovascularization in a mouse model

Subconjunctival gene delivery of the transcription factor GA-binding protein delays corneal neovascularization in a mouse model

Tumor Suppressor INK4: Determination of the Solution Structure of p18^INK4C and Demonstration of the Functional Significance of Loops in p18^INK4C and p16^INK4A ^,

Synergistic Action of GA-Binding Protein and Glucocorticoid Receptor in Transcription from the Mouse Mammary Tumor Virus Promoter

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The Structure of GABPα/β: An ETS Domain- Ankyrin Repeat Heterodimer Bound to DNA

Cited by 278 publications

References 26 publications

Subconjunctival gene delivery of the transcription factor GA-binding protein delays corneal neovascularization in a mouse model

Subconjunctival gene delivery of the transcription factor GA-binding protein delays corneal neovascularization in a mouse model

Tumor Suppressor INK4: Determination of the Solution Structure of p18INK4C and Demonstration of the Functional Significance of Loops in p18INK4C and p16INK4A ,

Synergistic Action of GA-Binding Protein and Glucocorticoid Receptor in Transcription from the Mouse Mammary Tumor Virus Promoter

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Tumor Suppressor INK4: Determination of the Solution Structure of p18^INK4C and Demonstration of the Functional Significance of Loops in p18^INK4C and p16^INK4A ^,