THE UNUSUAL STABILITY OF THE IS10 ANTI-SENSE RNA IS CRITICAL FOR ITS FUNCTION AND IS DETERMINED BY THE STRUCTURE OF ITS STEM-DOMAIN
THE UNUSUAL STABILITY OF THE IS10 ANTI-SENSE RNA IS CRITICAL FOR ITS FUNCTION AND IS DETERMINED BY THE STRUCTURE OF ITS STEM-DOMAIN
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DOI：
10.1002/j.1460-2075.1989.tb08616.x
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发表时间：
1989-12-20
期刊：
影响因子：
11.4
通讯作者：
SIMONS, RW
中科院分区：
文献类型：
--
作者：
CASE, CC;ROELS, SM;SIMONS, RW
IS 10转座由一个. apprx. 70 nt反义RNA，RNA-OUT。RNA-OUT折叠成双链体“茎域”，顶端是松散配对的“环域”。环结构域本身对于RNA-RNA配对至关重要;配对通过RNA-OUT环与靶mRNA的5“末端的相互作用而启动。我们在这里表明，RNA-OUT在体内是非常稳定的（半衰期60分钟），这种稳定性是由RNA-OUT干域的特定功能。一个关键特征是稳定的碱基配对：破坏茎配对的突变使体内RNA-OUT不稳定并废除反义控制;恢复配对的突变组合也恢复稳定性和控制。我们认为茎使RNA-OUT对3“核糖核酸外切酶具有抗性。茎域的其他特征阻止了这种必需的双链体成为双链核酸酶的有效底物：两个单碱基突变通过使RNA-OUT对RNA酶III敏感来破坏反义控制。 环区的突变对RNA-OUT稳定性几乎没有影响。IS 10生物学和有效的反义RNA的设计的影响进行了讨论。
IS10 transposition is regulated by an .apprx. 70 nt anti-sense RNA, RNA-OUT. RNA-OUT folds into a duplex ''stem-domain'' topped by a loosely paired ''loop-domain''. The loop-domain is critical for RNA-RNA pairing per se; pairing initiates by interaction of the RNA-OUT loop with the 5'' end of the target mRNA. We show here that RNA-OUT is unusually stable in vivo (half-life 60 min) and that this stability is conferred by specific features of the RNA-OUT stem-domain. One critical feature is stable base-pairing: mutations that disrupt stem pairing destabilize RNA-OUT in vivo and abolish anti-sense control; combinations of mutations that restore pairing also restore both stability and control. We propose that the stem renders RNA-OUT resistant to 3'' exoribonucleases. Other features of the stem-domain prevent this essential duplex from being an effective substrate for double-strand nucleases: two single base mutations disrupt antisense control by making RNA-OUT susceptible to RNase III. Mutations in the loop region have little effect on RNA-OUT stability. Implications for IS10 biology and the design of efficient anti-sense RNAs are discussed.