Shimizu T, Iwasato T, Yamagishi H
Department of Biophysics, Faculty of Science, Kyoto University, Japan.
J Exp Med. 1991 May 1;173(5):1065-72. doi: 10.1084/jem.173.5.1065.
We have identified circular DNAs containing the kappa light chain constant region (C kappa), as well as the excision products of V kappa-J kappa and V lambda-J lambda joining in adult mouse splenocytes. Analysis of C kappa-positive circular DNA clones revealed two recombination sites (intron recombining sequence [IRS]1 and -2) within the germline J kappa-C kappa intron region and the recombining sequence (RS) located downstream of the C kappa exon. While RS contains a conserved heptamer and nonamer separated by a 23-bp spacer on the 5' side, IRS1 sequence is an isolated heptamer without an obvious nonamer, and IRS2 contains a variant heptamer, CACAAAA. Since IRS1 and IRS2 recombined with both RS (23-bp spacer signal) and V kappa (12-bp spacer signal) with significant frequency, intron recombination sites seem to have dual recombination signals. These findings provide direct evidence that C kappa deletion preceding lambda gene rearrangement can occur by looping out and excision. Increased accessibility of inefficient recombinational loci within the intron may enable recombinase to accept wide signal sequence variation.
我们在成年小鼠脾细胞中鉴定出了包含κ轻链恒定区(Cκ)的环状DNA,以及Vκ-Jκ和Vλ-Jλ连接的切除产物。对Cκ阳性环状DNA克隆的分析揭示了种系Jκ-Cκ内含子区域内的两个重组位点(内含子重组序列[IRS]1和-2)以及位于Cκ外显子下游的重组序列(RS)。虽然RS在5'侧包含一个由23个碱基对间隔区隔开的保守七聚体和九聚体,但IRS1序列是一个孤立的七聚体,没有明显的九聚体,而IRS2包含一个变体七聚体CACAAAA。由于IRS1和IRS2与RS(23个碱基对间隔区信号)和Vκ(12个碱基对间隔区信号)都有显著频率的重组,内含子重组位点似乎具有双重重组信号。这些发现提供了直接证据,表明在λ基因重排之前,Cκ缺失可通过环出和切除发生。内含子内低效重组位点可及性的增加可能使重组酶能够接受广泛的信号序列变异。