Moore M W, Durdik J, Persiani D M, Selsing E
Proc Natl Acad Sci U S A. 1985 Sep;82(18):6211-5. doi: 10.1073/pnas.82.18.6211.
We isolated and characterized the germ-line counterpart of a DNA segment designated RS (for recombining sequence), that is frequently recombined in mouse lambda light chain-producing B lymphocytes. Using Southern blot analyses of myelomas and mouse-Chinese hamster fusion cell lines, we found that RS DNA sequences are located on mouse chromosome 6, evidently more than 15 kilobases downstream of the kappa light-chain locus. We find that a typical recognition site for Ig gene recombination is situated within germ-line RS sequences near the recombination points observed in at least two lambda chain-producing cell lines. This represents a complete and functional Ig recognition site that is not directly associated with Ig genes. We also characterized a recombined RS segment isolated from the cell line BM18-4.13.9. This recombined segment has a variable region kappa light chain gene (V kappa) joined directly to RS sequences. Our results suggest that the deletion of the kappa light chain constant region (C kappa) exon in many lambda chain-producing B cells is the result of RS recombination and that C kappa deletion may be mediated by the same processes as antibody gene V-J joining (J = joining segment gene). We discuss the potential biological significance of RS DNA recombination in B-cell maturation.
我们分离并鉴定了一个名为RS(重组序列)的DNA片段的种系对应物,该片段在产生小鼠λ轻链的B淋巴细胞中频繁发生重组。通过对骨髓瘤和小鼠-中国仓鼠融合细胞系进行Southern印迹分析,我们发现RS DNA序列位于小鼠6号染色体上,明显在κ轻链基因座下游超过15千碱基处。我们发现,在至少两个产生λ链的细胞系中观察到的重组点附近,Ig基因重组的典型识别位点位于种系RS序列内。这代表了一个完整且有功能的Ig识别位点,它不与Ig基因直接相关。我们还对从细胞系BM18 - 4.13.9中分离出的重组RS片段进行了鉴定。这个重组片段有一个可变区κ轻链基因(Vκ)直接与RS序列相连。我们的结果表明,许多产生λ链的B细胞中κ轻链恒定区(Cκ)外显子的缺失是RS重组的结果,并且Cκ缺失可能由与抗体基因V - J连接(J = 连接段基因)相同的过程介导。我们讨论了RS DNA重组在B细胞成熟中的潜在生物学意义。