Mizuguchi G, Nakagoshi H, Nagase T, Nomura N, Date T, Ueno Y, Ishii S
Tsukuba Life Science Center, Institute of Physical and Chemical Research, Ibaraki, Japan.
J Biol Chem. 1990 Jun 5;265(16):9280-4.
Three members of the myb gene family have been identified in human cDNA libraries c-myb, A-myb, and B-myb. We compared the DNA binding properties of the B-myb and c-myb proteins (B-MYB and c-MYB) using bacterially synthesized B-MYB and c-MYB in DNase I footprinting. B-MYB bound to most of the c-MYB binding sites examined, including the c-MYB binding site, MBS-I, in the simian virus (SV) 40 enhancer, in which the most frequent sequence was CCTAACTG. The MBS-I site was an enhancer element dependent on B-MYB and c-MYB in a co-transfection assay that used the B-myb or c-myb expression plasmid. Some sites in the SV40 genome, including the MBS-BI site, had high affinity with B-MYB but little or no affinity with c-MYB, in which the most frequent sequence was AGAAANPyrG. The MBS-BI site was an enhancer element dependent on B-MYB and a very weakly dependent on c-MYB. Our results showed that B-MYB is a transcriptional activator, like c-MYB, and that although B-MYB and c-MYB have similar sequence specificity for DNA binding some sequences were recognized by B-MYB preferentially.
在人类cDNA文库中已鉴定出myb基因家族的三个成员——c-myb、A-myb和B-myb。我们使用细菌合成的B-MYB和c-MYB,通过DNase I足迹法比较了B-myb和c-myb蛋白(B-MYB和c-MYB)的DNA结合特性。B-MYB与大多数检测的c-MYB结合位点结合,包括猿猴病毒(SV)40增强子中的c-MYB结合位点MBS-I,其最常见的序列是CCTAACTG。在使用B-myb或c-myb表达质粒的共转染实验中,MBS-I位点是一个依赖于B-MYB和c-MYB的增强子元件。SV40基因组中的一些位点,包括MBS-BI位点,与B-MYB具有高亲和力,但与c-MYB的亲和力很小或没有,其最常见的序列是AGAAANPyrG。MBS-BI位点是一个依赖于B-MYB的增强子元件,对c-MYB的依赖性非常弱。我们的结果表明,B-MYB与c-MYB一样,是一种转录激活因子,并且尽管B-MYB和c-MYB对DNA结合具有相似的序列特异性,但有些序列优先被B-MYB识别。