Shaffer A L, Peng A, Schlissel M S
Department of Molecular Biology and Genetics, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Immunity. 1997 Feb;6(2):131-43. doi: 10.1016/s1074-7613(00)80420-3.
The immunoglobulin kappa light chain locus has two enhancer elements: the intronic enhancer, which lies between the Jkappa cluster and the Ckappa exon, and the 3'kappa enhancer, which is located downstream of Ckappa. To address the contribution of these elements to the developmentally regulated activation of germline kappa locus transcription and rearrangement, we purified primary pro- and pre-B cells and determined by in vivo footprinting the sites within each enhancer that were occupied. We found that the kappa intronic enhancer NF-kappaB site is occupied in both pro- and pre-B cells, while CRE, BSAP, and PU.1/pip sites within the 3'kappa enhancer undergo changes in occupancy as cells progress from the pro-B to the pre-B cell stage. These findings suggest that regulation of the kappa locus in primary pre-B cells may be mediated by factors that bind the 3'kappa enhancer.
免疫球蛋白κ轻链基因座有两个增强子元件:内含子增强子,位于Jκ簇和Cκ外显子之间;3'κ增强子,位于Cκ下游。为了研究这些元件对种系κ基因座转录和重排的发育调控激活的贡献,我们纯化了原B细胞和前B细胞,并通过体内足迹法确定每个增强子内被占据的位点。我们发现,κ内含子增强子的NF-κB位点在原B细胞和前B细胞中均被占据,而随着细胞从原B细胞阶段发展到前B细胞阶段,3'κ增强子内的CRE、BSAP和PU.1/pip位点的占据情况发生了变化。这些发现表明,原前B细胞中κ基因座的调控可能由与3'κ增强子结合的因子介导。