Bulfone-Paus S, Reiners-Schramm L, Lauster R
Institute for Immunology, Freie Universität Berlin, Germany.
Nucleic Acids Res. 1995 Jun 11;23(11):1997-2005. doi: 10.1093/nar/23.11.1997.
Characterization of the regulatory elements involved in V(D)J recombination is crucial for understanding development of the B and T cell immune repertoire. Previously we have shown that the chicken immunoglobulin lambda light chain gene (CLLCG) undergoes lymphoid-specific rearrangement in transgenic mice. The whole gene is only 10 kb in length and contains all phylogenetically conserved target sites for recombinational and transcriptional regulation. In this study we have localized an enhancer element in a region 4 kb downstream of the constant (C) region. The 467 bp element can be subdivided into three subfragments. The previously detected silencer element on the V-J intervening sequence is shown to be localized on a 500 bp fragment. Partial silencer activity is retained on a 250 bp fragment, which includes an octamer motif. By mutational analysis this octamer is shown to be essential for B cell- but not for T cell-specific silencer function. The silencer represses transcription directed by heterologous elements like the SV 40 promoter or the Ig kappa 3' enhancer. We propose that transcription of the unrearranged and rearranged Ig genes is regulated by complex interactions between different modules from the promoter, enhancer and silencer, which is eliminated by recombination during B cell development.
鉴定参与V(D)J重排的调控元件对于理解B细胞和T细胞免疫库的发育至关重要。此前我们已经表明,鸡免疫球蛋白λ轻链基因(CLLCG)在转基因小鼠中发生淋巴细胞特异性重排。整个基因长度仅为10 kb,包含所有系统发育保守的重组和转录调控靶位点。在本研究中,我们在恒定(C)区下游4 kb的区域定位了一个增强子元件。这个467 bp的元件可细分为三个亚片段。先前在V-J间隔序列上检测到的沉默子元件位于一个500 bp的片段上。在一个包含八聚体基序的250 bp片段上保留了部分沉默子活性。通过突变分析表明,这个八聚体对于B细胞特异性沉默子功能是必需的,但对于T细胞特异性沉默子功能不是必需的。该沉默子抑制由异源元件如SV 40启动子或Igκ 3'增强子指导的转录。我们提出,未重排和重排的Ig基因的转录受来自启动子、增强子和沉默子的不同模块之间复杂相互作用的调控,这种调控在B细胞发育过程中通过重组被消除。