Department of Clinical Laboratory, Third Affiliated Hospital of Third Military Medical University, Chongqing, 400042, China.
Mol Biol Rep. 2010 Dec;37(8):3747-55. doi: 10.1007/s11033-010-0028-z. Epub 2010 Mar 26.
B-lymphocyte-induced maturation protein1(Blimp-1) and B-cell maturation antigen (BCMA) are essential factors in the development and survival of plasma cells. However, whether Blimp-1 could regulate the expression of BCMA is unknown. We found that the BCMA promoter region did not have typical "TATA" and "CAAT" box, but contained several potential binding sites of transcription factors, including the consensus sequences for Blimp-1, located in the "-31 to -21" and "-46 to -36" from the potential transcription initiation site of the mouse BCMA gene, respectively. Furthermore, induction of Blimp-1 over-expression significantly up-regulated the expression of BCMA and increased the BCMA promoter activity in mouse J558L plasma cells. In parallel, knockdown of Blimp-1 expression by the Blimp-1-specific shRNA significantly reduced the BCMA mRNA transcription and protein expression in J558L cells in vitro. Substitution mutation of the "-38 to -42" sequence, but not the "-23 to -27", in the BCMA promoter abolished the regulatory effect of Blimp-1 on the expression of BCMA. Importantly, Blimp-1 bound to the "GAAAC", but not its mutant "GATTC", contained BCMA promoter, as determined by competitive electrophoretic mobility shift assay (EMSA). Therefore, our data clearly suggest that Blimp-s a positive regulator of the expression of BCMA gene in mouse plasma cells.
B 淋巴细胞诱导成熟蛋白 1(Blimp-1)和 B 细胞成熟抗原(BCMA)是浆细胞发育和存活的重要因素。然而,Blimp-1 是否能调节 BCMA 的表达尚不清楚。我们发现,BCMA 启动子区域没有典型的“TATA”和“CAAT”框,但包含几个潜在的转录因子结合位点,包括 Blimp-1 的共有序列,分别位于潜在的转录起始位点的-31 到-21 和-46 到-36。此外,诱导 Blimp-1 过表达显著上调了 BCMA 的表达,并增加了小鼠 J558L 浆细胞中 BCMA 启动子的活性。同时,通过 Blimp-1 特异性 shRNA 敲低 Blimp-1 表达,显著降低了 J558L 细胞中 BCMA 的 mRNA 转录和蛋白表达。BCMA 启动子中“-38 到-42”序列的取代突变,但不是“-23 到-27”,消除了 Blimp-1 对 BCMA 表达的调节作用。重要的是,通过竞争性电泳迁移率变动分析(EMSA)确定,Blimp-1 结合到“GAAAC”,而不是其突变体“GATTC”,含有 BCMA 启动子。因此,我们的数据清楚地表明,Blimp-1 是小鼠浆细胞中 BCMA 基因表达的正调节因子。