Differential binding of nuclear factors to the intron 1 sequences containing the transcriptional pause site...

The molecular mechanisms that modulate c-myb mRNA levels in hematopoietic cells appear to involve premature termination of transcription in the first intron of the gene. We have examined the DNA-protein interactions within the first intron of the c-myb gene and identified a 1.0-kilobase region that could be responsible for its transcriptional regulation. Using the mobility-shift assay, we show a direct correlation between the extent of sequence-specific protein binding to intron 1 DNA fragments, and c-myb mRNA levels in different cell types. During dimethyl sulfoxide-induced differentiation of mouse erythroleukemic cells, there was a dramatic decrease in these nuclear factors that correlated with the decrease in the levels of c-myb mRNA. Nucleotide sequence analysis and DNase I footprinting revealed the presence of putative regulatory elements that are implicated in the binding of these nuclear factors. We propose that binding of nuclear factors to the site of transcriptional pause could play an important role in the regulation of c-myb transcription.