Brown B D, Harland R M
Division of Biochemistry and Molecular Biology, University of California, Berkeley 94720.
Genes Dev. 1990 Nov;4(11):1925-35. doi: 10.1101/gad.4.11.1925.
We have identified a messenger RNA (mRNA) sequence from a Xenopus homeo box-containing gene that is the target for a sequence-specific endoribonuclease in vivo. Synthetic RNA transcribed from an allele of the maternal gene Xlhbox2B is efficiently cleaved when injected into Xenopus oocytes. The cleavage sequence lies between the protein-coding region and a 600-base 3'-untranslated region. Intermediates in degradation are readily observed: Both the 5' and 3' products of cleavage are recovered, thus showing that the cleavage activity is an endonuclease. When a 90-base region of the Xlhbox2B sequence is inserted into a second homeo box RNA that is normally stable, it is sufficient to confer an identical cleavage reaction on the hybrid RNA. The cleaved region contains a repeated sequence motif and is cut at multiple sites. Inhibition of translation does not affect the rate or extent of cleavage, while the coinjection of antisense RNA complementary to the 90-base region completely blocks the reaction. Because most mRNAs are not found on polysomes during oogenesis, translation-independent cleavage at such sites may provide a novel post-transcriptional mechanism to regulate the amount of mRNA available for embryogenesis.
我们从非洲爪蟾一个含同源异型框的基因中鉴定出一种信使核糖核酸(mRNA)序列,它是体内一种序列特异性核糖核酸内切酶的作用靶点。当将从母源基因Xlhbox2B的一个等位基因转录而来的合成RNA注射到非洲爪蟾卵母细胞中时,它会被高效切割。切割序列位于蛋白质编码区和一个600个碱基的3'非翻译区之间。降解过程中的中间体很容易被观察到:切割产生的5'和3'产物都能被回收,这表明切割活性是由一种内切酶引起的。当将Xlhbox2B序列的一个90个碱基的区域插入到另一个通常稳定的同源异型框RNA中时,这足以使杂合RNA发生相同的切割反应。被切割的区域包含一个重复序列基序,并且在多个位点被切割。翻译抑制并不影响切割的速率或程度,而与90个碱基区域互补的反义RNA的共注射则完全阻断了该反应。由于在卵子发生过程中大多数mRNA并不存在于多核糖体上,在这些位点进行的不依赖翻译的切割可能提供一种新的转录后机制来调节可用于胚胎发育的mRNA量。