Naumovski L, Cleary M L
Department of Pediatrics, Division of Hematology/Oncology, Stanford University School of Medicine, California 94305, USA.
Mol Cell Biol. 1996 Jul;16(7):3884-92. doi: 10.1128/MCB.16.7.3884.
Using the yeast two-hybrid system, we have isolated a cDNA (designated BBP, for Bcl2-binding protein) for a protein (Bbp) that interacts with Bcl2. Bbp is identical to 53BP2, a partial clone of which was previously isolated in a two-hybrid screen for proteins that interact with p53. In this study, we show that specific interactions of Bbp/53BP2 with either Bcl2 or p53 require its ankyrin repeats and SH3 domain. These interactions can be reproduced in vitro with bacterially expressed fusion proteins, and competition experiments indicate that Bcl2 prevents p53 from binding to Bbp/53BP2. BBP/53BP2 mRNA is abundant in most cell lines examined, but the protein cannot be stably expressed in a variety of cell types by transfection. In transiently transfected cells, Bbp partially colocalizes with Bcl2 in the cytoplasm and results in an increased number of cells at G2/M, possibly accounting for the inability to obtain stable transfectants expressing the protein. These results demonstrate that a single protein can interact with either Bcl2 or p53 both in yeast cells and in vitro. The in vivo significance of these interactions and their potential consequences for cell cycle progression and cell death remain to be determined.
利用酵母双杂交系统,我们分离出了一个与Bcl2相互作用的蛋白质(Bbp)的cDNA(命名为BBP,即Bcl2结合蛋白)。Bbp与53BP2相同,53BP2的一个部分克隆先前在与p53相互作用的蛋白质的双杂交筛选中被分离出来。在本研究中,我们表明Bbp/53BP2与Bcl2或p53的特异性相互作用需要其锚蛋白重复序列和SH3结构域。这些相互作用可以在体外通过细菌表达的融合蛋白重现,竞争实验表明Bcl2阻止p53与Bbp/53BP2结合。BBP/53BP2 mRNA在大多数检测的细胞系中丰富,但通过转染该蛋白不能在多种细胞类型中稳定表达。在瞬时转染的细胞中,Bbp在细胞质中与Bcl2部分共定位,并导致G2/M期细胞数量增加,这可能解释了无法获得稳定表达该蛋白的转染子的原因。这些结果表明,单一蛋白质在酵母细胞和体外都能与Bcl2或p53相互作用。这些相互作用在体内的意义及其对细胞周期进程和细胞死亡的潜在影响仍有待确定。