Non-canonical 3'-5' extension of RNA with prebiotically plausible ribonucleoside 2',3'-cyclic phosphates.

Non-canonical 3'-5' extension of RNA with prebiotically plausible ribonucleoside 2',3'-cyclic phosphates.
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DOI:
10.1021/ja4127714
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发表时间:
2014-04-09
影响因子:
15
通讯作者:
Holliger P
Holliger P
中科院分区:
化学1区
文献类型:
--
作者:
Mutschler H;Holliger P

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核糖核苷2′,3 ′-环磷酸(N>p's)是由多种前生物学上合理的过程产生的,并且是早期RNA寡聚体组装的可靠构件。虽然N>p可以通过具有可变效率和区域选择性的非酶促过程聚合成短RNA,但是还没有描述用于RNA合成的酶促途径。在这里,我们报告这样一个非典型的3′-5′核苷酸转移酶活性。我们设计了一种发夹核酶的变体,用于催化所有四种N>p(2′,3 ′-环状A-、G-、U-和CMP)添加到RNA链的5′-羟基末端,在所有情况下,5′核苷酸的添加都通过在−7 °C下形成共晶冰相来增强。我们还观察到2′,3 ′-环磷酸激活的β-烟酰胺腺嘌呤二核苷酸(NAD>p)和ACA>p RNA三核苷酸的5′加成,以及GUCCA>p RNA五聚体的多次加成。我们的研究结果建立了一种新的RNA 3′-5′延伸模式,并对从益生元核苷酸库合成RNA寡聚体产生了影响。
Ribonucleoside 2′,3′-cyclic phosphates (N>p’s) are generated by multiple prebiotically plausible processes and are credible building blocks for the assembly of early RNA oligomers. While N>p’s can be polymerized into short RNAs by non-enzymatic processes with variable efficiency and regioselectivity, no enzymatic route for RNA synthesis had been described. Here we report such a non-canonical 3′-5′ nucleotidyl transferase activity. We engineered a variant of the hairpin ribozyme to catalyze addition of all four N>p’s (2′,3′-cyclic A-, G-, U-, and CMP) to the 5′-hydroxyl termini of RNA strands with 5′ nucleotide addition enhanced in all cases by eutectic ice phase formation at −7 °C. We also observed 5′ addition of 2′,3′-cyclic phosphate-activated β-nicotinamide adenine dinucleotide (NAD>p) and ACA>p RNA trinucleotide, and multiple additions of GUCCA>p RNA pentamers. Our results establish a new mode of RNA 3′-5′ extension with implications for RNA oligomer synthesis from prebiotic nucleotide pools.
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