Chen H, Zhang P, Radomska H S, Hetherington C J, Zhang D E, Tenen D G
Hematology/Oncology Division, Department of Medicine, Beth Israel Hospital, Harvard Medical School, Boston, Massachusetts 02215, USA.
J Biol Chem. 1996 Jun 28;271(26):15743-52. doi: 10.1074/jbc.271.26.15743.
PU.1 (spi-1), a member of the Ets transcription factor family, is predominantly expressed in myeloid and B cells, activates many B cell and myeloid genes, and is critical for development of both of these lineages. Our previous studies (Chen, H. M., Ray-Gallet, D., Zhang, P., Hetherington, C. J., Gonzalez, D. A., Zhang, D.-E., Moreau-Gachelin, F., and Tenen, D. G. (1995) Oncogene 11, 1549-1560) demonstrate that the PU.1 promoter directs cell type-specific reporter gene expression in myeloid cell lines, and that PU.1 activates its own promoter in an autoregulatory loop. Here we show that the murine PU.1 promoter is also specifically and highly functional in B cell lines as well. Oct-1 and Oct-2 can bind specifically to a site at base pair -55 in vitro, and this site is specifically protected in B cells in vivo. We also demonstrate that two other sites contribute to promoter activity in B cells; an Sp1 binding site adjacent to the octamer site, and the PU.1 autoregulatory site. Finally, we show that the B cell coactivator OBF-1/Bob1/OCA-B is only expressed in B cells and not in myeloid cells, and that OBF-1/Bob1/OCA-B can transactivate the PU.1 promoter in HeLa and myeloid cells. This B cell restricted coactivator may be responsible for the B cell specific expression of PU.1 mediated by the octamer site.
PU.1(spi-1)是Ets转录因子家族的成员,主要在髓系细胞和B细胞中表达,可激活许多B细胞和髓系细胞基因,对这两个细胞谱系的发育至关重要。我们之前的研究(Chen, H. M., Ray-Gallet, D., Zhang, P., Hetherington, C. J., Gonzalez, D. A., Zhang, D.-E., Moreau-Gachelin, F., and Tenen, D. G. (1995) Oncogene 11, 1549 - 1560)表明,PU.1启动子在髓系细胞系中指导细胞类型特异性报告基因的表达,并且PU.1在一个自调节环中激活其自身的启动子。在这里我们表明,小鼠PU.1启动子在B细胞系中同样具有特异性且高度功能性。Oct-1和Oct-2在体外可特异性结合到位于碱基对-55处的位点,并且该位点在体内的B细胞中受到特异性保护。我们还证明另外两个位点对B细胞中的启动子活性有贡献;一个与八聚体位点相邻的Sp1结合位点以及PU.1自调节位点。最后,我们表明B细胞共激活因子OBF-1/Bob1/OCA-B仅在B细胞中表达而不在髓系细胞中表达,并且OBF-1/Bob1/OCA-B可在HeLa细胞和髓系细胞中转激活PU.1启动子。这种B细胞限制性共激活因子可能负责由八聚体位点介导的PU.1的B细胞特异性表达。