Characterization of the sequence-specific interaction of mouse c-myb protein with DNA.

摘要: We have examined parameters that affect sequence-specific interactions of the mouse c-myb protein with DNA oligomers containing Myb-binding motif (CA/CGTTPu). Complexes formed between these and in vitro translated proteins were analysed by electrophoresis on non-denaturing polyacrylamide gels using mobility-shift assay. By progressive truncation coding sequences it was demonstrated amino acids downstream a region three imperfect 51-52 residue repeats (designated R1, R2 R3), which are located close to terminus protein, had no qualitative or quantitative effect ability interact specifically this motif. However, removal only five R3 repeat completely abolished activity. The contribution individual DNA-binding domain interaction investigated precisely deleting each individually: combination absolutely required for complex formation while R1 dispensible. showed quantitatively greater duplicated rather than single motif, particular where arranged tandem. Moreover, observed interacted tandem motifs as monomer. These findings imply subunit straddles adjacent binding sites implications activity discussed.