Kuwata S, Masuta C, Takanami Y
Life Science Research Laboratory, Japan Tobacco Incorporated, Yokohama.
J Gen Virol. 1991 Oct;72 ( Pt 10):2385-9. doi: 10.1099/0022-1317-72-10-2385.
Cucumber mosaic virus Y satellite RNA (Y-satRNA) induces distinctive yellow mosaic symptoms on tobacco, whereas S19 satellite RNA (S19-satRNA) causes an attenuated green mosaic on tobacco, although they show considerable sequence identity. Biological assays of infectious chimeric satellite RNA molecules synthesized from cDNA clones of Y-satRNA and S19-satRNA using common restriction sites showed that the determinant for the induction of yellow mosaic symptoms lies in the BstXI-NheI fragment, in which 14 nucleotide differences are found between the two satellite RNAs. To define more precisely the yellow mosaic determinant(s) in this fragment, several site-directed mutants of Y-satRNA were created. The replacement of AUU, at nucleotides 191 to 193 in Y-satRNA, with GC, which mimics the S19-satRNA sequence at the corresponding site, abolished the ability of Y-satRNA to elicit a yellow mosaic. Conversely, a mutant RNA molecule derived from S19-satRNA in which GC at nucleotides 192 and 193 was changed to AUU induced the yellow mosaic symptoms. Thus, the phenotypes of two satellite RNAs on tobacco can be altered reciprocally by changing the sequences in this limited region.
黄瓜花叶病毒Y卫星RNA(Y-satRNA)在烟草上诱导出独特的黄色花叶症状,而S19卫星RNA(S19-satRNA)在烟草上引起减弱的绿色花叶症状,尽管它们显示出相当高的序列同一性。使用常见限制位点从Y-satRNA和S19-satRNA的cDNA克隆合成的感染性嵌合卫星RNA分子的生物学分析表明,诱导黄色花叶症状的决定因素位于BstXI-NheI片段中,在这两个卫星RNA之间发现了14个核苷酸差异。为了更精确地定义该片段中的黄色花叶决定因素,创建了几个Y-satRNA的定点突变体。将Y-satRNA中第191至193位核苷酸的AUU替换为GC,这模拟了S19-satRNA在相应位点的序列,消除了Y-satRNA引发黄色花叶的能力。相反,一个源自S19-satRNA的突变RNA分子,其中第192和193位核苷酸的GC被改变为AUU,诱导了黄色花叶症状。因此,通过改变这个有限区域的序列,可以相互改变两种卫星RNA在烟草上的表型。