Motoyama N, Okada H, Azuma T
Department of Molecular Biology, Nagoya City University School of Medicine, Japan.
Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):7933-7. doi: 10.1073/pnas.88.18.7933.
To study the distribution of somatic mutation, we determined nucleotide sequences of rearranged lambda 1-chain genomic DNA from four hybridomas obtained from C57BL/6 mice that had been immunized with (4-hydroxy-3-nitrophenyl)acetyl-conjugated chicken gamma globulin. In total, 114 nucleotide substitutions were observed, with neither insertion nor deletion. Sixty-one mutations occurred in the variable-joining region genes (V lambda 1-J lambda 1) and 49 in joining-constant (J lambda 1-C lambda 1) introns. Although frequency decreased with distance from the V lambda 1-J lambda 1 coding region, somatic mutations occurred in the entire J lambda 1-C lambda 1 intron and even in the C lambda 1 region. We found four nucleotide substitutions in C lambda 1 genes, all of which were replacement mutations. Therefore, the mechanism responsible for somatic mutation is operative into the C lambda 1 exons. Nucleotide sequences of rearranged but inactive lambda 2-chain genes from two hybridomas were also examined and compared with those of lambda 1-chain genes. The clustering of replacement mutations in complementarity-determining regions in the inactive lambda 2-chain genes similar to the active lambda 1-chain genes suggested a mechanism that induces somatic mutation preferentially in this region even in the absence of antigenic selection.
为了研究体细胞突变的分布情况,我们测定了从四只经(4-羟基-3-硝基苯基)乙酰共轭鸡γ球蛋白免疫的C57BL/6小鼠获得的杂交瘤中重排的λ1链基因组DNA的核苷酸序列。总共观察到114个核苷酸替换,无插入或缺失。61个突变发生在可变连接区基因(Vλ1-Jλ1),49个发生在连接恒定区(Jλ1-Cλ1)内含子中。尽管频率随着与Vλ1-Jλ1编码区距离的增加而降低,但体细胞突变发生在整个Jλ1-Cλ1内含子甚至Cλ1区域。我们在Cλ1基因中发现了四个核苷酸替换,均为置换突变。因此,负责体细胞突变的机制在Cλ1外显子中起作用。还检查了来自两个杂交瘤的重排但无活性的λ2链基因的核苷酸序列,并与λ1链基因的序列进行了比较。无活性λ2链基因中互补决定区置换突变的聚集类似于活性λ1链基因,这表明即使在没有抗原选择的情况下,也存在一种机制优先在该区域诱导体细胞突变。