B-cell- and myocyte-specific E2-box-binding factors contain E12/E47-like subunits

Recent studies have identified a family of DNA-binding proteins that share a common DNA-binding and dimerization domain with the potential to form a helix-loop-helix (HLH) structure. Various HLH proteins can form heterodimers that bind to a common DNA sequence, termed the E2-box. We demonstrate here that E2-box-binding B-cell- and myocyte-specific nuclear factors contain subunits which are identical or closely related to ubiquitously expressed (E12/E47) HLH proteins. These biochemical function for E12/E47-like molecules in mammalian differentiation, similar to the genetically defined function of daughterless in Drosophila development.

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Indirect Effects of DNA Sequence on Transcriptional Activation by Prokaryotic DNA Binding Proteins

Mechanisms of Transcription - Nucleic Acids and Molecular Biology ◽

10.1007/978-3-642-60691-5_9 ◽

1997 ◽

pp. 115-134

Author(s):

S. Hilchey ◽

J. Xu ◽

G. B. Koudelka

Keyword(s):

Dna Binding ◽

Dna Sequence ◽

Transcriptional Activation ◽

Indirect Effects ◽

Binding Proteins ◽

Dna Binding Proteins

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The protein Id: A negative regulator of helix-loop-helix DNA binding proteins

Cell ◽

10.1016/0092-8674(90)90214-y ◽

1990 ◽

Vol 61 (1) ◽

pp. 49-59 ◽

Cited By ~ 1357

Author(s):

Robert Benezra ◽

Robert L. Davis ◽

Daniel Lockshon ◽

David L. Turner ◽

Harold Weintraub

Keyword(s):

Dna Binding ◽

Binding Proteins ◽

Dna Binding Proteins ◽

Negative Regulator ◽

Helix Loop Helix

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Built by association: structure and function of helix-loop-helix DNA-binding proteins

Structure ◽

10.1016/s0969-2126(00)00002-2 ◽

1994 ◽

Vol 2 (1) ◽

pp. 1-4 ◽

Cited By ~ 16

Author(s):

Simon EV Phillips

Keyword(s):

Dna Binding ◽

Binding Proteins ◽

Dna Binding Proteins ◽

Structure And Function ◽

Helix Loop Helix ◽

Association Structure ◽

And Function

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Expression of ID, a negative regulator of helix-loop-helix DNA binding proteins, is down-regulated at confluence and enhanced by dexamethasone in a mouse osteoblastic cell line, MC3T3E1

Biochemical and Biophysical Research Communications ◽

10.1016/s0006-291x(05)81322-1 ◽

1991 ◽

Vol 180 (3) ◽

pp. 1194-1199 ◽

Cited By ~ 26

Author(s):

Toshiko Ogata ◽

Masaki Noda

Keyword(s):

Dna Binding ◽

Cell Line ◽

Binding Proteins ◽

Dna Binding Proteins ◽

Negative Regulator ◽

Osteoblastic Cell ◽

Osteoblastic Cell Line ◽

Helix Loop Helix

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E2A and E2-2 are subunits of B-cell-specific E2-box DNA-binding proteins

Molecular and Cellular Biology ◽

10.1128/mcb.13.6.3522-3529.1993 ◽

1993 ◽

Vol 13 (6) ◽

pp. 3522-3529

Author(s):

G Bain ◽

S Gruenwald ◽

C Murre

Keyword(s):

Monoclonal Antibodies ◽

B Cells ◽

Dna Binding ◽

B Cell ◽

Dna Sequences ◽

Dna Binding Proteins ◽

Gene Products ◽

Helix Loop Helix ◽

The Difference

A class of helix-loop-helix (HLH) proteins, including E2A (E12 and E47), E2-2, and HEB, that bind in vitro to DNA sequences present in the immunoglobulin (Ig) enhancers has recently been identified. E12, E47, E2-2, and HEB are each present in B cells. The presence of many different HLH proteins raises the question of which of the HLH proteins actually binds the Ig enhancer elements in B cells. Using monoclonal antibodies specific for both E2A and E2-2, we show that both E2-2 and E2A polypeptides are present in B-cell-specific Ig enhancer-binding complexes. E2-box-binding complexes in pre-B cells contain both E2-2 and E2A HLH subunits, whereas in mature B cells only E2A gene products are present. We show that the difference in E2-box-binding complexes in pre-B and mature B cells may be caused by differential expression of E2A and E2-2.

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eyelid antagonizes wingless signaling during Drosophila development and has homology to the Bright family of DNA-binding proteins

Genes & Development ◽

10.1101/gad.11.15.1949 ◽

1997 ◽

Vol 11 (15) ◽

pp. 1949-1962 ◽

Cited By ~ 61

Author(s):

J. E. Treisman ◽

A. Luk ◽

G. M. Rubin ◽

U. Heberlein

Keyword(s):

Dna Binding ◽

Binding Proteins ◽

Dna Binding Proteins ◽

Drosophila Development ◽

Wingless Signaling

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Fission Yeast Homologs of Human CENP-B Have Redundant Functions Affecting Cell Growth and Chromosome Segregation

Molecular and Cellular Biology ◽

10.1128/mcb.20.8.2852-2864.2000 ◽

2000 ◽

Vol 20 (8) ◽

pp. 2852-2864 ◽

Cited By ~ 37

Author(s):

Mary Baum ◽

Louise Clarke

Keyword(s):

Dna Binding ◽

Fission Yeast ◽

Dna Sequence ◽

Chromosome Segregation ◽

Binding Proteins ◽

Dna Binding Proteins ◽

Central Core ◽

Cell Extracts ◽

Redundant Functions

ABSTRACT Two functionally important DNA sequence elements in centromeres of the fission yeast Schizosaccharomyces pombe are the centromeric central core and the K-type repeat. Both of these DNA elements show internal functional redundancy that is not correlated with a conserved DNA sequence. Specific, but degenerate, sequences in these elements are bound in vitro by the S. pombeDNA-binding proteins Abp1p (also called Cbp1p) and Cbhp, which are related to the mammalian centromere DNA-binding protein CENP-B. In this study, we determined that Abp1p binds to at least one of its target sequences within S. pombe centromere II central core (cc2) DNA with an affinity (Ks = 7 × 109 M−1) higher than those of other known centromere DNA-binding proteins for their cognate targets. In vivo, epitope-tagged Cbhp associated with centromeric K repeat chromatin, as well as with noncentromeric regions. Likeabp1+/cbp1 +, we found thatcbh + is not essential in fission yeast, but a strain carrying deletions of both genes (Δabp1 Δcbh) is extremely compromised in growth rate and morphology and missegregates chromosomes at very high frequency. The synergism between the two null mutations suggests that these proteins perform redundant functions in S. pombe chromosome segregation. In vitro assays with cell extracts with these proteins depleted allowed the specific assignments of several binding sites for them within cc2 and the K-type repeat. Redundancy observed at the centromere DNA level appears to be reflected at the protein level, as no single member of the CENP-B-related protein family is essential for proper chromosome segregation in fission yeast. The relevance of these findings to mammalian centromeres is discussed.

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Novel method for identifying sequence-specific DNA-binding proteins.

Molecular and Cellular Biology ◽

10.1128/mcb.5.9.2307 ◽

1985 ◽

Vol 5 (9) ◽

pp. 2307-2315 ◽

Cited By ~ 22

Author(s):

D Levens ◽

P M Howley

Keyword(s):

Dna Binding ◽

Fusion Protein ◽

Dna Sequence ◽

Binding Proteins ◽

Binding Protein ◽

Protein S ◽

Dna Binding Proteins ◽

Lac Repressor ◽

Lac Operator ◽

Beta Galactosidase

We developed a general method for the enrichment and identification of sequence-specific DNA-binding proteins. A well-characterized protein-DNA interaction is used to isolate from crude cellular extracts or fractions thereof proteins which bind to specific DNA sequences; the method is based solely on this binding property of the proteins. The DNA sequence of interest, cloned adjacent to the lac operator DNA segment is incubated with a lac repressor-beta-galactosidase fusion protein which retains full operator and inducer binding properties. The DNA fragment bound to the lac repressor-beta-galactosidase fusion protein is precipitated by the addition of affinity-purified anti-beta-galactosidase immobilized on beads. This forms an affinity matrix for any proteins which might interact specifically with the DNA sequence cloned adjacent to the lac operator. When incubated with cellular extracts in the presence of excess competitor DNA, any protein(s) which specifically binds to the cloned DNA sequence of interest can be cleanly precipitated. When isopropyl-beta-D-thiogalactopyranoside is added, the lac repressor releases the bound DNA, and thus the protein-DNA complex consisting of the specific restriction fragment and any specific binding protein(s) is released, permitting the identification of the protein by standard biochemical techniques. We demonstrate the utility of this method with the lambda repressor, another well-characterized DNA-binding protein, as a model. In addition, with crude preparations of the yeast mitochondrial RNA polymerase, we identified a 70,000-molecular-weight peptide which binds specifically to the promoter region of the yeast mitochondrial 14S rRNA gene.

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