Johnson D G, Carayannopoulos L, Capra J D, Tucker P W, Hanke J H
Department of Microbiology, Southwestern Medical Center, Dallas, Texas 75235-9096.
Mol Cell Biol. 1990 Mar;10(3):982-90. doi: 10.1128/mcb.10.3.982-990.1990.
All immunoglobulin genes contain a conserved octanucleotide promoter element, ATGCAAAT, which has been shown to be required for their normal B-cell-specific transcription. Proteins that bind this octamer have been purified, and cDNAs encoding octamer-binding proteins have been cloned. Some of these proteins (referred to as OTF-2) are lymphoid specific, whereas at least one other, and possibly more (referred to as OTF-1), is found ubiquitously in all cell types. The exact role of these different proteins in directing the tissue-specific expression of immunoglobulin genes is unclear. We have identified two human pre-B-cell lines that contain extremely low levels of OTF-2 yet still express high levels of steady-state immunoglobulin heavy-chain mRNA in vivo and efficiently transcribe an immunoglobulin gene in vitro. Addition of a highly enriched preparation of OTF-1 made from one of these pre-B cells or from HeLa cells specifically stimulated in vitro transcription of an immunoglobulin gene. Furthermore, OFT-1 appeared to have approximately the same transactivation ability as OTF-2 when normalized for binding activity. These results suggest that OTF-1, without OTF-2, is sufficient for transcription of immunoglobulin genes and that OTF-2 alone is not responsible for the B-cell-specific regulation of immunoglobulin gene expression.
所有免疫球蛋白基因都含有一个保守的八核苷酸启动子元件ATGCAAAT,该元件已被证明是其正常B细胞特异性转录所必需的。与该八聚体结合的蛋白质已被纯化,编码八聚体结合蛋白的cDNA也已被克隆。其中一些蛋白质(称为OTF-2)是淋巴细胞特异性的,而至少还有一种(可能更多,称为OTF-1)在所有细胞类型中普遍存在。这些不同蛋白质在指导免疫球蛋白基因的组织特异性表达中的确切作用尚不清楚。我们鉴定出两个人类前B细胞系,它们含有极低水平的OTF-2,但在体内仍表达高水平的稳态免疫球蛋白重链mRNA,并在体外有效转录免疫球蛋白基因。添加由其中一个前B细胞或经体外特异性刺激的HeLa细胞制备的高度富集的OTF-1制剂,可特异性刺激免疫球蛋白基因的体外转录。此外,当以结合活性进行标准化时,OTF-1似乎具有与OTF-2大致相同的反式激活能力。这些结果表明,没有OTF-2时,OTF-1足以进行免疫球蛋白基因的转录,并且单独的OTF-2并不负责免疫球蛋白基因表达的B细胞特异性调节。