A conserved motif of vertebrate Y RNAs essential for chromosomal DNA replication
A conserved motif of vertebrate Y RNAs essential for chromosomal DNA replication
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DOI：
10.1261/rna.1472009
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发表时间：
2009-07-01
期刊：
影响因子：
4.5
通讯作者：
Krude, Torsten
中科院分区：
文献类型：
--
作者：
Gardiner, Timothy J.;Christov, Christo P.;Krude, Torsten
非编码Y RNA是人类无细胞系统中G1期晚期模板细胞核中染色体DNA复制重建所必需的。Y RNA基因存在于所有脊椎动物和一些孤立的非脊椎动物中，但Y RNA功能的保守性及其功能的关键决定因素尚不清楚。在这里，我们确定了一个决定Y RNA功能的DNA复制，这是保守的整个脊椎动物进化。脊椎动物Y RNA能够在人类无细胞DNA复制系统中重建染色体DNA复制，但非脊椎动物Y RNA不能。脊椎动物Y RNA双链茎中的保守核苷酸序列基序与Y RNA功能相关。人Y1 RNA突变体的功能筛选鉴定了这种保守的基序作为体外重建DNA复制的重要决定因素。包含该RNA基序的双链RNA寡核苷酸足以重建DNA复制，但相应的DNA或随机序列RNA寡核苷酸则不能。在完整的细胞中，野生型hY1或保守的RNA双链体可以拯救对hY3 RNA的RNA干扰后的DNA复制抑制。因此，我们已经确定了一个新的RNA基序，它在脊椎动物Y RNA进化中是保守的，并且是人类染色体DNA复制中Y RNA功能所必需的和足够的。
Noncoding Y RNAs are required for the reconstitution of chromosomal DNA replication in late G1 phase template nuclei in a human cell-free system. Y RNA genes are present in all vertebrates and in some isolated nonvertebrates, but the conservation of Y RNA function and key determinants for its function are unknown. Here, we identify a determinant of Y RNA function in DNA replication, which is conserved throughout vertebrate evolution. Vertebrate Y RNAs are able to reconstitute chromosomal DNA replication in the human cell-free DNA replication system, but nonvertebrate Y RNAs are not. A conserved nucleotide sequence motif in the double-stranded stem of vertebrate Y RNAs correlates with Y RNA function. A functional screen of human Y1 RNA mutants identified this conserved motif as an essential determinant for reconstituting DNA replication in vitro. Double-stranded RNA oligonucleotides comprising this RNA motif are sufficient to reconstitute DNA replication, but corresponding DNA or random sequence RNA oligonucleotides are not. In intact cells, wild-type hY1 or the conserved RNA duplex can rescue an inhibition of DNA replication after RNA interference against hY3 RNA. Therefore, we have identified a new RNA motif that is conserved in vertebrate Y RNA evolution, and essential and sufficient for Y RNA function in human chromosomal DNA replication.