Mouse RSS spacer sequences affect the rate of V(D)J recombination
Mouse RSS spacer sequences affect the rate of V(D)J recombination
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DOI：
10.1007/bf02660064
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发表时间：
1996-01-01
期刊：
影响因子：
3.2
通讯作者：
Wu, GE
中科院分区：
文献类型：
--
作者：
Fanning, L;Connor, A;Wu, GE
翻译信号序列（RSS）是参与V（D）J重组的位点。RSS由被12或23 bp的间隔区隔开的7和9 bp基序组成（分别为RSS-12和RSS-23）。先前已经报道，间隔区的DNA序列在RSS重组中不起作用。使用灵敏的染色体外V（D）J重组试验重新检查了这一发现。通过瞬时转染将RSS的组织和序列不同的纯化底物引入Abelson小鼠白血病病毒转化的小鼠细胞系中，并测量RSS的使用率。比较两种基材：两种底物都含有串联的2 Igk-V12-RSS和Igk-J123-RSS，但一种底物（DR 92）含有2 Igk-V12-RSS的天然序列（CTACAGACTGGA），而另一种底物（DR 93）在近端Igk-V12-RSS的间隔区（CTACCGACTGGA）中含有C5而不是A5。DR 93的RSSV（D）J重组率低于DR 92（54 vs. 37%;P= 0.0024）。结果表明，RSS间隔区序列影响V（D）J重组率。
Recombination signal sequences (RSS) are the sites involved inV(D)Jrecombination. RSS consist of 7 and 9 bp motifs separated by a spacer of 12 or 23 bp (RSS-12 and RSS-23 respectively). It has been previously reported that the DNA sequence of the spacers does not play a role in RSS recombination. This finding was re-examined using a sensitive extrachromosomalV(D)Jrecombination assay. Recombination substrates differing in organization and sequences of the RSS were introduced by transient transfection into an Abelson mouse leukaemia virus-transformed mouse cell line and the rate of usage of RSS was measured. Two substrates were compared: both had 2Igk-V12-RSS in tandem and anIgk-J123-RSS, but one substrate (DR92) had the natural sequence for the 2Igk-V12-RSS (CTACAGACTGGA), while the other (DR93) had a C5instead of an A5in the spacer of the proximalIgk-V12-RSS (CTACCGACTGGA). The rate of RSSV(D)Jrecombination was lower in DR93 than in DR92 (54 vs. 37%;P= 0.0024). The results indicate that RSS spacer sequence affects the rate ofV(D)Jrecombination.