Kemler I, Bucher E, Seipel K, Müller-Immerglück M M, Schaffner W
Institut für Molekularbiologie II, Universität Zürich, Switzerland.
Nucleic Acids Res. 1991 Jan 25;19(2):237-42. doi: 10.1093/nar/19.2.237.
Immunoglobulin (Ig) gene promoters contain the octamer sequence motif ATGCAAAT which is recognized by cellular transcription factors (Oct factors). Besides the ubiquitous Oct-1 factor, there is also a group of related factors (Oct-2 factors) encoded by a separate gene. The Oct-2 gene is regulated in a cell-type specific manner, and the protein is present in large amounts in B lymphocytes. We have previously shown that simple composite promoters of an octamer/TATA box type are poorly active in non-B cells but are strongly responsive to ectopic expression of Oct-2A factor, a major representative of the lymphocyte Oct-2 factors. In the present study we have tested the activity of a number of composite promoters and natural Ig promoters, and their response to Oct-1 and Oct-2 factors. Unexpectedly, we find that octamer/TATA promoters with a high affinity octamer site direct ubiquitous expression. By contrast, promoter constructions that behave in a B cell-specific manner tend to have a weak octamer binding site. These promoters are responsive to ectopic expression of additional Oct-factor, irrespective of whether it is Oct-1 or Oct-2. Using natural Ig promoters rather than composite promoters, we find that an IgH promoter is well transcribed in non-B cells via the ubiquitous Oct-1 factor, while Ig kappa and Ig lambda light chain promoters require additional Oct factor for maximal expression. It seems therefore likely that during B cell differentiation, Ig heavy chain promoters can be activated by Oct-1, before the appearance of Oct-2 factors. Oct-2 factors then would serve to boost the expression from Ig light chain promoters, which are known to be activated only after successful heavy chain gene rearrangement.
免疫球蛋白(Ig)基因启动子包含八聚体序列基序ATGCAAAT,该基序可被细胞转录因子(Oct因子)识别。除了普遍存在的Oct-1因子外,还有一组由单独基因编码的相关因子(Oct-2因子)。Oct-2基因以细胞类型特异性方式受到调控,并且该蛋白在B淋巴细胞中大量存在。我们之前已经表明,八聚体/TATA盒类型的简单复合启动子在非B细胞中活性较差,但对淋巴细胞Oct-2因子的主要代表Oct-2A因子的异位表达有强烈反应。在本研究中,我们测试了许多复合启动子和天然Ig启动子的活性,以及它们对Oct-1和Oct-2因子的反应。出乎意料的是,我们发现具有高亲和力八聚体位点的八聚体/TATA启动子指导普遍表达。相比之下,表现出B细胞特异性的启动子构建体往往具有较弱的八聚体结合位点。这些启动子对额外Oct因子的异位表达有反应,无论它是Oct-1还是Oct-2。使用天然Ig启动子而非复合启动子,我们发现IgH启动子通过普遍存在的Oct-1因子在非B细胞中能很好地转录,而Igκ和Igλ轻链启动子需要额外的Oct因子才能实现最大表达。因此,在B细胞分化过程中,Ig重链启动子似乎可以在Oct-2因子出现之前被Oct-1激活。然后Oct-2因子将有助于增强Ig轻链启动子的表达,已知这些轻链启动子仅在重链基因成功重排后才被激活。