Fukuhara T, Moriyama H, Nitta T
Laboratory of Molecular Cell Biology, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Japan.
J Biol Chem. 1995 Jul 28;270(30):18147-9. doi: 10.1074/jbc.270.30.18147.
A linear, plasmid-like, double-stranded RNA (dsRNA) was isolated from rice, and its entire sequence of 13,952 nucleotides (nt) was determined. The dsRNA encodes a single, unusually long, open reading frame (13,716 nt, 4,572 amino acid residues), which includes an RNA helicase-like domain and an RNA-dependent RNA polymerase-like domain. A series of Northern hybridization and primer extension experiments revealed that the coding (sense) strand of the dsRNA contains a discontinuity (nick) at a position 1,211 nt (or 1,256 nt) from the 5' end. This discontinuity divides not only the coding strand of dsRNA molecule into a 1,211-nt fragment and a 12,741-nt fragment (or a 1,256-nt fragment and a 12,696-nt fragment) but also divides the long open reading frame into a 5' part of 1,045 nt (348 amino acid residues) and a 3' part of 12,671 nt (4,224 amino acid residues) or a 5' part of 1,090 nt (363 amino acid residues) and a 3' part of 12,626 nt (4,209 amino acid residues). It seems likely that almost all dsRNA molecules in rice plants contain such a discontinuity. This rice dsRNA appears to be a novel and unique RNA replicon.
从水稻中分离出一种线性的、类似质粒的双链RNA(dsRNA),并测定了其13952个核苷酸(nt)的完整序列。该dsRNA编码一个单一的、异常长的开放阅读框(13716 nt,4572个氨基酸残基),其中包括一个RNA解旋酶样结构域和一个RNA依赖性RNA聚合酶样结构域。一系列Northern杂交和引物延伸实验表明,dsRNA的编码(正义)链在距5'端1211 nt(或1256 nt)处存在一个间断(切口)。这种间断不仅将dsRNA分子的编码链分为一个1211 nt的片段和一个12741 nt的片段(或一个1256 nt的片段和一个12696 nt的片段),还将长开放阅读框分为一个1045 nt(348个氨基酸残基)的5'部分和一个12671 nt(4224个氨基酸残基)的3'部分,或一个1090 nt(363个氨基酸残基)的5'部分和一个12626 nt(4209个氨基酸残基)的3'部分。水稻植株中几乎所有的dsRNA分子似乎都含有这样的间断。这种水稻dsRNA似乎是一种新颖独特的RNA复制子。