Perry R P, Kelley D E, Schibler U
Proc Natl Acad Sci U S A. 1979 Aug;76(8):3678-82. doi: 10.1073/pnas.76.8.3678.
To help elucidate the mechanisms of formation and expression of active immunoglobulin genes, we have made a systematic study of the processing patterns of the mRNAs that code for a closely related family of kappa chains. Among the members of this family, represented by the V(kappa)21 group of plasmacytomas, are examples in which six different germ-line V genes are joined to four different J segments in various combinations. The mRNA precursors were identified by hybridizing a cloned kappa-cDNA probe to poly(A)-containing nuclear RNAs that were size fractionated on methylmercury-agarose gels and transferred to diazotized paper. Based on the length of the segment excised in the last detectable processing step, which presumably represents the removal of the J-C intron, the precursor patterns were classified into four primary categories that correlated well with the type of J segment being expressed. The J segments were thus located at distances ranging from 2.4 to 4.8 kilobases from the constant-region gene. Different V genes joined to the same J segment exhibited similar precursor patterns, suggesting that the size of the translocated V segment may be relatively uniform among members of the V(kappa)21 group. A large (9.1 kilobases) component, which is likely to represent a primary transcription product, was observed in all processing categories, regardless of which J segment was being utilized. This surprising observation leads to some interesting predictions about the mechanism of V-J translocation.
为了帮助阐明活性免疫球蛋白基因的形成和表达机制,我们对编码密切相关的κ链家族的mRNA加工模式进行了系统研究。在以浆细胞瘤的V(κ)21组为代表的该家族成员中,存在六个不同的种系V基因以各种组合与四个不同的J片段连接的例子。通过将克隆的κ-cDNA探针与含poly(A)的核RNA杂交来鉴定mRNA前体,这些核RNA在甲基汞-琼脂糖凝胶上进行大小分级,然后转移到重氮化纸上。根据在最后可检测到的加工步骤中切除的片段长度(推测代表J-C内含子的去除),前体模式被分为四个主要类别,这与所表达的J片段类型密切相关。因此,J片段位于距恒定区基因2.4至4.8千碱基的距离范围内。连接到相同J片段的不同V基因表现出相似的前体模式,这表明在V(κ)21组成员中,易位的V片段大小可能相对一致。在所有加工类别中都观察到一个大的(9.1千碱基)成分,它可能代表初级转录产物,无论使用哪个J片段。这一惊人的观察结果对V-J易位机制产生了一些有趣的预测。