Sunter G, Bisaro D M
Department of Molecular Genetics, Ohio State University, Columbus 13210.
Virology. 1989 Dec;173(2):647-55. doi: 10.1016/0042-6822(89)90577-1.
In a previous study, the bipartite genome of tomato golden mosaic virus (TGMV) was shown to be transcribed into at least six polyadenylated RNAs (G. Sunter, W.E. Gardiner, and D. M. Bisaro, 1989, Virology 170, 243-250). Two of these, a 1.3-kb complementary sense and a 0.9-kb viral sense transcript, were mapped to the B genome component of this geminivirus. The results of more detailed primer extension and S1 nuclease protection experiments presented here define the limits of the single transcription unit corresponding to the 0.9-kb RNA which spans the BR1 open reading frame (ORF). The data also demonstrate that complementary sense TGMV RNAs are more complex than indicated by our earlier studies. Analysis of the 1.3-kb BL1-specific RNA indicates that it is actually a family of distinct transcripts with different start sites. Three transcripts have 5' ends that map near the common region of DNA B and all of these start sites lie upstream of the BL1 ORF. Similar analysis of the 1.6-kb complementary sense AL1 RNA indicates that a complex set of transcripts also map to the analogous region of genome component A. Four transcripts have 5' ends that map near the common region but only one of these start sites is upstream of the initiation codon for the AL1 open reading frame (ORF). None of the transcripts appear to be processed. The possible significance of multiple transcripts in these regions of the TGMV genome is discussed, and the common region-proximal transcription units of the A and B genome components are compared.
在先前的一项研究中,番茄金色花叶病毒(TGMV)的二分基因组被证明转录为至少六种多聚腺苷酸化RNA(G. 桑特、W.E. 加德纳和D.M. 比萨罗,1989年,《病毒学》170卷,243 - 250页)。其中两种,一种1.3 kb的互补义转录本和一种0.9 kb的病毒义转录本,被定位到这种双生病毒的B基因组组分上。此处呈现的更详细的引物延伸和S1核酸酶保护实验结果确定了对应于跨越BR1开放阅读框(ORF)的0.9 kb RNA的单个转录单元的界限。数据还表明,TGMV互补义RNA比我们早期研究显示的更为复杂。对1.3 kb BL1特异性RNA的分析表明,它实际上是一个具有不同起始位点的不同转录本家族。三种转录本的5'端定位在DNA B的共同区域附近,并且所有这些起始位点都位于BL1 ORF的上游。对1.6 kb互补义AL1 RNA的类似分析表明,一组复杂的转录本也定位到基因组组分A的类似区域。四种转录本的5'端定位在共同区域附近,但这些起始位点中只有一个位于AL1开放阅读框(ORF)起始密码子的上游。没有一种转录本似乎经过加工。讨论了TGMV基因组这些区域中多个转录本的可能意义,并比较了A和B基因组组分的共同区域近端转录单元。