BOG pRb Binding Protein for Cell-cycle Control/transform

BOG(RBBP-9)是一种新的细胞蛋白，通过LXCXE基序与pRb、p107和p130相互作用。纯合的bog-/-小鼠已经产生，在纯合的动物中没有检测到bog mRNA和蛋白的水平。沼泽-/-出生时健康，与沼泽+/-和野生型动物相比，生长速度正常，并具有生育能力。目前，我们正在对沼泽-/-鼠的大量苏木精和曙红染色组织进行组织学评估。我们已经开始使用从13.5天的沼泽/胚胎分离的小鼠胚胎成纤维细胞(MEF)，按照标准程序(3T3和3T9方案)寻找BOG在细胞周期控制中的作用，并用于体外实验。初步结果表明，与野生型(WT)细胞相比，BOG-/-MEF的增殖能力更为有限。-/-细胞比WT更早停止增殖，导致细胞总数缓慢但逐渐减少。在高密度和低密度条件下，结果是一致的。以WST-1为基础，用两个新品系重复进行增殖试验。结果证实，BOG-/-MEF的生长能力受损与该基因有直接关系。结合所获得的数据和增殖分析，表明沼泽缺失的MEF表现出“提前衰老”的表型。为了验证这一假设，我们选择了衰老相关的β-半乳糖苷酶活性作为衰老标记。这一结果与沼泽缺陷细胞早期获得的复制衰老状态是一致的。

BOG (RBBP-9) is a novel cellular protein which interacts with pRb, p107 and p130 through the LXCXE motif. The homozygous BOG-/- mouse has been generated, and no detectible level of BOG mRNA nor protein was found in the homozygous animals. BOG-/- are born healthy, grow at a normal rate compared with BOG+/- and wildtype animals, and are fertile. At present we are performing histological evaluations of numerous haematoxylin and eosin stained tissues from BOG-/- mice. We have started to use mouse embryo fibroblasts (MEFs) isolated from 13.5 day old BOG -/- embryos in search of a role for BOG in cell cycle control, following standard procedures (3T3 and 3T9 protocols) and used for in vitro experiments. Preliminary results indicate that BOG-/- MEFs possess a more limited capacity to proliferate than the wildtype (WT) cells. The -/- cells stopped proliferating sooner than the WT resulting in a slow but progressive decrease in the total number of cells. The results were identical under both high and low cell density conditions. The WST-1 based proliferation assay was repeated using two new lines. The results confirm that the impared growth capacity of BOG-/- MEF is directly related to the genotype. Taking together the data obtained with the proliferation assays suggested that BOG null MEFs exhibited a "premature senescence" phenotype. To test that hypothesis, we chose the senescence associated b-galactosidase activity as a senescence marker. The results were consistent with an earlier acquisition of a replicative senescence status in the BOG deficient cells.