Schubart D B, Sauter P, Massa S, Friedl E M, Schwarzenbach H, Matthias P
Friedrich Miescher Institute, Basel, Switzerland.
Nucleic Acids Res. 1996 May 15;24(10):1913-20. doi: 10.1093/nar/24.10.1913.
The B cell-specific activity of immunoglobulin (Ig) gene promoters is to a large extent mediated by the conserved octamer motif ATTTGCAT. This requires the DNA binding octamer factors Oct-1 and/or Oct-2, as well as an additional B cell-restricted non-DNA binding cofactor. We recently cloned such a coactivator specific for Oct-1 or Oct-2 from human B cells and called it OBF-1. Here we report the isolation and characterization of the murine homologue. Full-length cDNA clones as well as genomic clones were isolated and the gene structure was determined. The deduced protein sequence shows that the mouse protein has an identical length, is likewise proline rich and shows 89% overall identity to the human protein. The OBF-1 gene is expressed in a very highly B cell-specific manner and is transcribed in cells representative of all stages of B cell differentiation, including the earliest ones. We show that OBF-1 interacts in the absence of DNA with the POU domain of Oct-1 or Oct-2 and also with the general transcription factors TBP and TFIIB. Furthermore, we demonstrate that although OBF-1 efficiently activates promoter octamer sites, it does not activate enhancer octamer sites.
免疫球蛋白(Ig)基因启动子的B细胞特异性活性在很大程度上由保守的八聚体基序ATTTGCAT介导。这需要DNA结合八聚体因子Oct-1和/或Oct-2,以及另外一种B细胞限制性非DNA结合辅因子。我们最近从人B细胞中克隆了这种对Oct-1或Oct-2特异的共激活因子,并将其命名为OBF-1。在此我们报道小鼠同源物的分离和鉴定。分离出了全长cDNA克隆以及基因组克隆,并确定了基因结构。推导的蛋白质序列表明,小鼠蛋白长度相同,同样富含脯氨酸,与人类蛋白的总体一致性为89%。OBF-1基因以非常高的B细胞特异性方式表达,并且在B细胞分化所有阶段的代表性细胞中都有转录,包括最早阶段的细胞。我们表明,OBF-1在无DNA的情况下与Oct-1或Oct-2的POU结构域相互作用,还与通用转录因子TBP和TFIIB相互作用。此外,我们证明,尽管OBF-1能有效激活启动子八聚体位点,但不能激活增强子八聚体位点。