Lavender P, Vandel L, Bannister A J, Kouzarides T
Wellcome/CRC Institute and Department of Pathology, University of Cambridge, UK.
Oncogene. 1997 Jun 5;14(22):2721-8. doi: 10.1038/sj.onc.1201243.
A yeast two-hybrid screen has identified HBP1 as a transcription factor capable of interacting with the pocket protein family. We show that HBP1 can interact with one of these, RB, both in vitro and in mammalian cells. Two distinct RB binding sites are present within HBP1--a high affinity binding site, mediated by an LXCXE motif and a separate low affinity binding site present within an activation domain. GAL4-fusion experiments indicate that HBP1 contains a masked activation domain. Deletion of two independent N- and C-terminal inhibitor domains unmasks an activation domain which is 100-fold more active than the full length protein. The released activation capacity is repressed by RB, p130 and p107. In addition, E1A can repress the activity of HBP1 via conserved region 1 sequences in a manner independent of the CBP co-activator. We show by stable expression in NIH3T3 cells that HBP1 has the capacity to induce morphological transformation of cells in culture.
酵母双杂交筛选已鉴定出HBP1是一种能够与口袋蛋白家族相互作用的转录因子。我们证明,HBP1在体外和哺乳动物细胞中均能与其中之一的RB相互作用。HBP1内存在两个不同的RB结合位点——一个由LXCXE基序介导的高亲和力结合位点,以及一个位于激活域内的单独的低亲和力结合位点。GAL4融合实验表明,HBP1含有一个被掩盖的激活域。删除两个独立的N端和C端抑制域可使激活域暴露,该激活域的活性比全长蛋白高100倍。释放的激活能力受到RB、p130和p107的抑制。此外,E1A可以通过保守区域1序列以独立于CBP共激活因子的方式抑制HBP1的活性。我们通过在NIH3T3细胞中的稳定表达表明,HBP1有能力诱导培养细胞的形态转化。