Degradation of nanoRNA is performed by multiple redundant RNases in Bacillus subtilis.

Degradation of nanoRNA is performed by multiple redundant RNases in Bacillus subtilis.
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DOI:
10.1093/nar/gkp527
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发表时间:
2009-08
影响因子:
14.9
通讯作者:
Mechold U
Mechold U
中科院分区:
生物学2区
文献类型:
--
作者:
Fang M;Zeisberg WM;Condon C;Ogryzko V;Danchin A;Mechold U

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大肠杆菌只有一种必需的寡核糖核酸酶(Orn),这种酶可以降解长度较短的5个残基的寡核糖核酸(nanoRNA)。包括枯草芽孢杆菌在内的厚壁菌门没有Orn同源物。我们之前已经在枯草芽孢杆菌中鉴定出YtqI (NrnA)是Orn的功能类似物。从枯草芽孢杆菌的基因组文库中筛选可以补充条件orn突变的基因,我们在这里确定了YngD (NrnB)作为枯草芽孢杆菌的第二个纳米ornase。与NrnA一样,NrnB也是磷酸酯酶DHH/DHHA1蛋白家族的成员。与Orn类似,NrnB在体外降解纳米rna 5-mers。NrnB的低表达水平足以实现orn的互补。YhaM是枯草芽孢杆菌中存在的一种已知的rna酶,在体外有效地降解nanoRNA,但需要高水平表达才能部分互补。枯草芽孢杆菌的三突变体(nrnA -, nrnB -, yhaM -)是有活力的,并且几乎没有显示出生长损伤。最后,RNase J1似乎对纳米RNA也有一些5 ‘到3 ’的外核糖核酸酶活性,因此可以潜在地完成RNA的降解。我们得出的结论是,与大肠杆菌不同,纳米rna的降解在枯草芽孢杆菌中以冗余的方式进行。
Escherichia coli possesses only one essential oligoribonuclease (Orn), an enzyme that can degrade oligoribonucleotides of five residues and shorter in length (nanoRNA). Firmicutes including Bacillus subtilis do not have an Orn homolog. We had previously identified YtqI (NrnA) as functional analog of Orn in B. subtilis. Screening a genomic library from B. subtilis for genes that can complement a conditional orn mutant, we identify here YngD (NrnB) as a second nanoRNase in B. subtilis. Like NrnA, NrnB is a member of the DHH/DHHA1 protein family of phosphoesterases. NrnB degrades nanoRNA 5-mers in vitro similarily to Orn. Low expression levels of NrnB are sufficient for orn complementation. YhaM, a known RNase present in B. subtilis, degrades nanoRNA efficiently in vitro but requires high levels of expression for only partial complementation of the orn– strain. A triple mutant (nrnA–, nrnB–, yhaM–) in B. subtilis is viable and shows almost no impairment in growth. Lastly, RNase J1 seems also to have some 5′-to-3′ exoribonuclease activity on nanoRNA and thus can potentially finish degradation of RNA. We conclude that, unlike in E. coli, degradation of nanoRNA is performed in a redundant fashion in B. subtilis.
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