Abstract
B-MYB is an ubiquitous protein required for mammalian cell growth. In this report we show that B-MYB transactivates its own promoter through a 120 bp segment proximal to the transcription start site. The B-MYB-responsive element does not contain myb-binding sites and gel-shift analysis shows that SP1, but not B-MYB, protein contained in SAOS2 cell extracts binds to the 120 bp B-myb promoter fragment. B-MYB-dependent transactivation is cooperatively increased in the presence of SP1, but not SP3 overexpression. When the SP1 elements of the B-myb promoter are transferred in front of a heterologous promoter, an increased response to B-MYB results. In contrast, c-MYB, the prototype member of the Myb family, is not able to activate the luciferase construct containing the SP1 elements. With the use of an SP1-GAL4 fusion protein, we have determined that the cooperative activation occurs through the domain A of SP1. These observations suggest that B-MYB functions as a coactivator of SP1, and that diverse combinations of myb and SP1 sites may dictate the responsiveness of myb-target genes to the various members of the myb family.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenovirus E1A Proteins / metabolism
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Binding Sites
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Cell Cycle Proteins*
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Cell Line
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DNA-Binding Proteins / genetics*
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DNA-Binding Proteins / metabolism
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Fungal Proteins / genetics
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Fungal Proteins / metabolism
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Herpes Simplex Virus Protein Vmw65 / metabolism
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Humans
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NF-kappa B / metabolism
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Oligonucleotides / pharmacology
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Promoter Regions, Genetic*
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Protein Binding
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Proto-Oncogene Proteins / metabolism
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Proto-Oncogene Proteins c-myb
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Recombinant Fusion Proteins
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Saccharomyces cerevisiae Proteins*
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Sp1 Transcription Factor / genetics
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Sp1 Transcription Factor / metabolism*
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Sp3 Transcription Factor
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Trans-Activators / genetics*
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Trans-Activators / metabolism
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Transcription Factors / genetics
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Transcription Factors / metabolism
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Transcription, Genetic
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Transcriptional Activation* / drug effects
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Transfection
Substances
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Adenovirus E1A Proteins
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Cell Cycle Proteins
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DNA-Binding Proteins
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Fungal Proteins
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GAL4 protein, S cerevisiae
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Herpes Simplex Virus Protein Vmw65
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MYBL2 protein, human
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NF-kappa B
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Oligonucleotides
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Proto-Oncogene Proteins
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Proto-Oncogene Proteins c-myb
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Recombinant Fusion Proteins
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SP3 protein, human
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Saccharomyces cerevisiae Proteins
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Sp1 Transcription Factor
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Trans-Activators
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Transcription Factors
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Sp3 Transcription Factor