Characterization of Spi-B, a transcription factor related to the putative oncoprotein Spi-1/PU.1.

We have cloned a human cDNA from a new gene, spi-B, on the basis of its homology with the DNA-binding domain of the Spi-1/PU.1 putative oncogene product. spi-B codes for a protein of 262 amino acids presenting 43% overall identity with Spi-1. Its highly basic carboxy-terminal region exhibits 34% sequence identity with the DNA-binding domain of the Ets-1 protein. We showed that the Spi-B protein is able to bind the purine-rich sequence (PU box) recognized by Spi-1/PU.1 and to activate transcription of a reporter plasmid containing PU boxes. Chromosome in situ hybridization allowed us to map spi-B to the 19q13.3-19q13.4 region of the human genome. spi-B, like spi-1, was found to be expressed in various murine and human hematopoietic cell lines except T lymphoid cell lines.

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The carboxy-terminal transactivation domain of Oct-4 acquires cell specificity through the POU domain.

Molecular and Cellular Biology ◽

10.1128/mcb.17.1.154 ◽

1997 ◽

Vol 17 (1) ◽

pp. 154-162 ◽

Cited By ~ 67

Author(s):

A Brehm ◽

K Ohbo ◽

H Schöler

Keyword(s):

Dna Binding ◽

Cell Lines ◽

Regulatory Mechanism ◽

Cell Types ◽

Binding Domain ◽

Dna Binding Domain ◽

Cell Type ◽

Pou Domain ◽

Cell Type Specific ◽

Carboxy Terminal

The POU transcription factor Oct-4 is expressed in totipotent and pluripotent cells of the early mouse embryo and the germ cell lineage. Transactivation capacities of regions flanking the DNA binding domain of Oct-4 were analyzed in undifferentiated and differentiated cell lines. The amino- and carboxy-terminal regions (N domain and C domain) fused to the Gal4 DNA binding domain both functioned as transactivation domains in all cell lines tested. However, the C domain failed to activate transcription in some cell lines in the context of the native protein. The underlying regulatory mechanism appears to involve the POU domain of Oct-4 and can discriminate between different POU domains, since constructs in which the C domain was instead fused to the POU domain of Pit-1 were again equally active in all cell lines. These results indicate that the C domain is subject to cell-type-specific regulation mediated by the Oct-4 POU domain. Phosphopeptide analysis revealed that the cell-type-specific difference of C-domain activity correlates with a difference in Oct-4 phosphorylation status. Since Oct-4 is expressed in a variety of distinct cell types during murine embryogenesis, these results suggest an additional regulatory mechanism for determining Oct-4 function in rapidly changing cell types during development.

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Transactivation Functions of the N-Terminal Domains of Nuclear Hormone Receptors: Protein Folding and Coactivator Interactions

Molecular Endocrinology ◽

10.1210/me.2002-0258 ◽

2003 ◽

Vol 17 (1) ◽

pp. 1-10 ◽

Cited By ~ 125

Author(s):

Raj Kumar ◽

E. Brad Thompson

Keyword(s):

Dna Binding ◽

Protein Interactions ◽

Hormone Receptors ◽

Nuclear Hormone Receptor ◽

Binding Domain ◽

Dna Binding Domain ◽

Protein Protein Interactions ◽

Carboxy Terminal ◽

And Function ◽

Terminal Domains

Abstract The N-terminal domains (NTDs) of many members of the nuclear hormone receptor (NHR) family contain potent transcription-activating functions (AFs). Knowledge of the mechanisms of action of the NTD AFs has lagged, compared with that concerning other important domains of the NHRs. In part, this is because the NTD AFs appear to be unfolded when expressed as recombinant proteins. Recent studies have begun to shed light on the structure and function of the NTD AFs. Recombinant NTD AFs can be made to fold by application of certain osmolytes or when expressed in conjunction with a DNA-binding domain by binding that DNA-binding domain to a DNA response element. The sequence of the DNA binding site may affect the functional state of the AFs domain. If properly folded, NTD AFs can bind certain cofactors and primary transcription factors. Through these, and/or by direct interactions, the NTD AFs may interact with the AF2 domain in the ligand binding, carboxy-terminal portion of the NHRs. We propose models for the folding of the NTD AFs and their protein-protein interactions.

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Characterization of the Elk-1 ETS DNA-binding Domain

Journal of Biological Chemistry ◽

10.1074/jbc.270.11.5805 ◽

1995 ◽

Vol 270 (11) ◽

pp. 5805-5811 ◽

Cited By ~ 32

Author(s):

Paul Shore ◽

Louise Bisset ◽

Jeremy Lakey ◽

Jonathan P. Waltho ◽

Richard Virden ◽

...

Keyword(s):

Dna Binding ◽

Binding Domain ◽

Dna Binding Domain

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Expression and Characterization of a Fusion Protein Between the Catalytic Domain of Poly(ADP-Ribose) polymerase and the DNA Binding Domain of the Glucocorticoid Receptor

Biochemical and Biophysical Research Communications ◽

10.1006/bbrc.1994.2012 ◽

1994 ◽

Vol 202 (2) ◽

pp. 880-887 ◽

Cited By ~ 2

Author(s):

D. Rosenthal ◽

T. Hong ◽

B. Cherney ◽

S.M. Zhang ◽

T. Shima ◽

...

Keyword(s):

Dna Binding ◽

Glucocorticoid Receptor ◽

Fusion Protein ◽

Catalytic Domain ◽

Binding Domain ◽

Dna Binding Domain

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Dissection of a carboxy-terminal region of the yeast regulatory protein RAP1 with effects on both transcriptional activation and silencing

Molecular and Cellular Biology ◽

10.1128/mcb.12.3.1209-1217.1992 ◽

1992 ◽

Vol 12 (3) ◽

pp. 1209-1217

Author(s):

C F Hardy ◽

D Balderes ◽

D Shore

Keyword(s):

Dna Binding ◽

Binding Site ◽

Transcriptional Activation ◽

Binding Sites ◽

Binding Protein ◽

Binding Domain ◽

Dominant Negative Effect ◽

Dna Binding Domain ◽

Wild Type ◽

Carboxy Terminal

RAP1 is an essential sequence-specific DNA-binding protein in Saccharomyces cerevisiae whose binding sites are found in a large number of promoters, where they function as upstream activation sites, and at the silencer elements of the HMR and HML mating-type loci, where they are important for repression. We have examined the involvement of specific regions of the RAP1 protein in both repression and activation of transcription by studying the properties of a series of hybrid proteins containing RAP1 sequences fused to the DNA-binding domain of the yeast protein GAL4 (amino acids 1 to 147). GAL4 DNA-binding domain/RAP1 hybrids containing only the carboxy-terminal third of the RAP1 protein (which lacks the RAP1 DNA-binding domain) function as transcriptional activators of a reporter gene containing upstream GAL4 binding sites. Expression of some hybrids from the strong ADH1 promoter on multicopy plasmids has a dominant negative effect on silencers, leading to either partial or complete derepression of normally silenced genes. The GAL4/RAP1 hybrids have different effects on wild-type and several mutated but functional silencers. Silencers lacking either an autonomously replicating sequence consensus element or the RAP1 binding site are strongly derepressed, whereas the wild-type silencer or a silencer containing a deletion of the binding site for another silencer-binding protein, ABF1, are only weakly affected by hybrid expression. By examining a series of GAL4 DNA-binding domain/RAP1 hybrids, we have mapped the transcriptional activation and derepression functions to specific parts of the RAP1 carboxy terminus.(ABSTRACT TRUNCATED AT 250 WORDS)

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Isolation and characterization of the core single-stranded DNA-binding domain of purine-rich element binding protein B (Purβ)

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2010.08.059 ◽

2010 ◽

Vol 400 (3) ◽

pp. 340-345 ◽

Cited By ~ 8

Author(s):

Amy E. Rumora ◽

Ashley N. Steere ◽

Jon E. Ramsey ◽

Anna M. Knapp ◽

Bryan A. Ballif ◽

...

Keyword(s):

Dna Binding ◽

Binding Protein ◽

Binding Domain ◽

Dna Binding Domain ◽

Single Stranded Dna ◽

The Core ◽

Isolation And Characterization ◽

Element Binding Protein ◽

Protein B

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Production and characterization of a retinoic acid receptor RARγ construction encompassing the DNA binding domain and the disordered N-terminal proline rich domain

Protein Expression and Purification ◽

10.1016/j.pep.2013.12.001 ◽

2014 ◽

Vol 95 ◽

pp. 113-120 ◽

Cited By ~ 3

Author(s):

Denise Martinez-Zapien ◽

Marc-André Delsuc ◽

Gilles Travé ◽

Régis Lutzing ◽

Cécile Rochette-Egly ◽

...

Keyword(s):

Retinoic Acid ◽

Dna Binding ◽

Retinoic Acid Receptor ◽

Binding Domain ◽

Dna Binding Domain ◽

Acid Receptor ◽

Rich Domain

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Dissection of a carboxy-terminal region of the yeast regulatory protein RAP1 with effects on both transcriptional activation and silencing.

Molecular and Cellular Biology ◽

10.1128/mcb.12.3.1209 ◽

1992 ◽

Vol 12 (3) ◽

pp. 1209-1217 ◽

Cited By ~ 49

Author(s):

C F Hardy ◽

D Balderes ◽

D Shore

Keyword(s):

Dna Binding ◽

Binding Site ◽

Transcriptional Activation ◽

Binding Sites ◽

Binding Protein ◽

Binding Domain ◽

Dominant Negative Effect ◽

Dna Binding Domain ◽

Wild Type ◽

Carboxy Terminal

RAP1 is an essential sequence-specific DNA-binding protein in Saccharomyces cerevisiae whose binding sites are found in a large number of promoters, where they function as upstream activation sites, and at the silencer elements of the HMR and HML mating-type loci, where they are important for repression. We have examined the involvement of specific regions of the RAP1 protein in both repression and activation of transcription by studying the properties of a series of hybrid proteins containing RAP1 sequences fused to the DNA-binding domain of the yeast protein GAL4 (amino acids 1 to 147). GAL4 DNA-binding domain/RAP1 hybrids containing only the carboxy-terminal third of the RAP1 protein (which lacks the RAP1 DNA-binding domain) function as transcriptional activators of a reporter gene containing upstream GAL4 binding sites. Expression of some hybrids from the strong ADH1 promoter on multicopy plasmids has a dominant negative effect on silencers, leading to either partial or complete derepression of normally silenced genes. The GAL4/RAP1 hybrids have different effects on wild-type and several mutated but functional silencers. Silencers lacking either an autonomously replicating sequence consensus element or the RAP1 binding site are strongly derepressed, whereas the wild-type silencer or a silencer containing a deletion of the binding site for another silencer-binding protein, ABF1, are only weakly affected by hybrid expression. By examining a series of GAL4 DNA-binding domain/RAP1 hybrids, we have mapped the transcriptional activation and derepression functions to specific parts of the RAP1 carboxy terminus.(ABSTRACT TRUNCATED AT 250 WORDS)

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