Biophysical characterization of the c-myb DNA-binding domain

Ebneth, Andreas; Schweers, O.; Thole, Hubert; Fagin, Ursula; Urbanke, Claus; Maass, Guenther; Wolfes, Heiner

doi:10.1021/bi00252a026

Cited by 17 publications

(12 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…EMSA experiments confirmed that at high concentrations bMYB305 indeed bound to mutated MRE cHs probes (data not shown). Similar observations have been made with cMYB (Ebneth et al, 1994).…”

Section: Pcmybi Interacts In Vivo With Mre Chssupporting

confidence: 88%

PcMYB1, a novel plant protein containing a DNA‐binding domain with one MYB repeat, interacts in vivo with a light‐regulatory promoter unit

et al. 1997

View full text Add to dashboard Cite

SummaryLight-regulatory unit 1 (LRU1) is necessary for and sufficient to mediate light-dependent activation of the chalcone synthase (CHS) minimal promoter in Petroselinum crispum. This composite promoter unit consists of at least two distinct c/s-acting elements, designated ACE ces and MRE cHs, both of which are required for light induction. The ACGT-containing element ACE cHs interacts with common plant regulatory factors (CPRFs) which belong to the basic region/leucine zipper (bZlP) class of transcription factors. Here, we demonstrate that MRE cHs, originally identified as an in vivo DNA footprint, is a MYB recognition element. This element possesses a functional core that is essential for light responsiveness and is specifically recognized by two distantly related MYB-like proteins: MYB305 and the novel factor MYB1 from P. crispum. PcMYB1 was identified by both its specific binding to MRE cHs in vitro and recognition of MRE cHs in vivo. The deduced amino acid sequence revealed that PcMYB1 contains only one MYB-like repeat. This portion of the protein constitutes the DNA-binding domain. Mutational analysis of PcMYB1 in combination with sequence comparison suggests the presence of a helix-turn-helix structure containing a recognition helix that is sufficient for sequence-specific binding. The structure of this distinct MYB-like DNA-binding domain appears to be conserved in proteins from all three eukaryotic phyla.

show abstract

“…EMSA experiments confirmed that at high concentrations bMYB305 indeed bound to mutated MRE cHs probes (data not shown). Similar observations have been made with cMYB (Ebneth et al, 1994).…”

Section: Pcmybi Interacts In Vivo With Mre Chssupporting

confidence: 88%

PcMYB1, a novel plant protein containing a DNA‐binding domain with one MYB repeat, interacts in vivo with a light‐regulatory promoter unit

et al. 1997

View full text Add to dashboard Cite

show abstract

“…The minimal region of c‐Myb that is both necessary and sufficient for sequence‐specific DNA‐binding has been narrowed down to repeats R2 and R3 [7,10–12] and unlike many other sequence‐specific DNA‐binding proteins, the Myb proteins appear to bind to DNA as monomers [10,11]. Though repeat R1 was not shown to be involved in sequence‐specific recognition, it has been suggested to increase the affinity of the protein for DNA and the stability of the Myb‐DNA complex [13–17].…”

Section: Introductionmentioning

confidence: 99%

Transcriptional activation by the Myb proteins requires a specific local promoter structure

1999

View full text Add to dashboard Cite

The biological effects of the cellular c-Myb and the viral v-Myb proteins are strikingly different. While c-Myb is indispensable for normal hematopoiesis, v-Myb induces acute leukemia. The v-Myb DNA-binding domain (DBD) differs from that of c-Myb mainly by deletion of the first of three repeats which correlates with efficient oncogenic transformation and a decrease in DNA-binding activity. To investigate the difference in DNA-binding and transcriptional activation, oligonucleotide selection and electrophoretic mobility shift assays were employed. The v-Myb DBD (R2R3) shows an intrinsic DNAbinding specificity for an AT-rich downstream extension of the Myb recognition element (MRE) PyAAC T / G G for efficient binding to this site, whereas R1 within the c-Myb DBD allows for more flexibility for this downstream extension. Therefore, due to the presence of repeat R1, c-Myb can bind to a greater number of target sites. The intrinsic DNA-binding specificity of R2R3 is further supported with the results from in vivo transcriptional activation experiments which demonstrated that both the v-Myb and c-Myb DBDs require an extension of the MRE (motif #1) by a downstream T-stretch (motif #2) for full activity. Surprisingly, the T-stretch improves binding when present on either strand, but is required on a specific strand for transcriptional activation.z 1999 Federation of European Biochemical Societies.

show abstract

“…The Myb DNA-binding domain comprises three imperfect tandem repeats (R1, R2, and R3), each forming a tri-helical protein fold in which an HTH motif is formed by helix 2 and 3 (32,33). Together, R2 and R3 are sufficient for recognition of specific DNA sequences, whereas R1 enhances the stability of the Myb-DNA complex (34,35). The three helices in each repeat are maintained by a hydrophobic core that includes three strictly conserved tryptophan residues ( Fig.…”

mentioning

confidence: 99%

Characterization of the Extended Myb-like DNA-binding Domain of Trithorax Group Protein Zeste

Mohrmann

Kal

Verrijzer

2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

Biophysical characterization of the c-myb DNA-binding domain

Cited by 17 publications

References 28 publications

PcMYB1, a novel plant protein containing a DNA‐binding domain with one MYB repeat, interacts in vivo with a light‐regulatory promoter unit

PcMYB1, a novel plant protein containing a DNA‐binding domain with one MYB repeat, interacts in vivo with a light‐regulatory promoter unit

Transcriptional activation by the Myb proteins requires a specific local promoter structure

Characterization of the Extended Myb-like DNA-binding Domain of Trithorax Group Protein Zeste

Contact Info

Product

Resources

About