Vera A, Daròs J A, Flores R, Hernández C
Instituto de Biología Molecular y Celular de Plantas, Universidad Politécnica de Valencia, 46022 Valencia, Spain.
J Virol. 2000 Nov;74(22):10390-400. doi: 10.1128/jvi.74.22.10390-10400.2000.
Carnation small viroid-like RNA (CarSV RNA) and its homologous DNA are the two forms of a unique plant retroviroid-like system. CarSV RNA is a 275-nucleotide noninfectious viroid-like RNA, present in certain carnation plants, which can adopt hammerhead structures in both polarity strands and self-cleave accordingly. CarSV DNA is organized as a series of head-to-tail multimers forming part of extrachromosomal elements in which CarSV DNA sequences are fused to sequences of carnation etched ring virus (CERV), a plant pararetrovirus. Analysis of more than 30 CarSV-CERV DNA junctions showed that distinct regions of the viral genome seem able to interact with CarSV DNA. All these junctions were short nucleotide stretches common to both CarSV and CERV DNAs. This suggests a polymerase-driven mechanism for their origin involving an enzyme with low processivity, most likely the viral reverse transcriptase. This view was further supported by the observation that most of CarSV sequences forming part of the junctions correspond either to strong secondary structure motifs in the conformation proposed for CarSV RNA or to its self-cleavage sites, which may have facilitated polymerase jumping. Accompanying the most-abundant CarSV RNA, a series of CarSV RNAs with sequence deletions were previously characterized. Here we have identified some of their corresponding DNA forms, together with other CarSV DNA forms with deletions not found in any CarSV RNA species identified so far. Some of these CarSV DNA forms have also been detected fused to CERV sequences. The existence of these shortened CarSV DNA versions may provide a continuous input of their corresponding transcripts and explain the persistence of CarSV RNAs with defective hammerhead structures for which an RNA-RNA model of amplification seems unlikely.
香石竹类病毒样RNA(CarSV RNA)及其同源DNA是一种独特的植物类逆转病毒样系统的两种形式。CarSV RNA是一种275个核苷酸的非感染性类病毒样RNA,存在于某些香石竹植物中,其两条极性链均可形成锤头结构并相应地进行自我切割。CarSV DNA被组织成一系列头尾相连的多聚体,构成染色体外元件的一部分,其中CarSV DNA序列与植物副逆转录病毒香石竹蚀刻环病毒(CERV)的序列融合。对30多个CarSV - CERV DNA连接点的分析表明,病毒基因组的不同区域似乎能够与CarSV DNA相互作用。所有这些连接点都是CarSV和CERV DNA共有的短核苷酸片段。这表明其起源涉及一种聚合酶驱动机制,该机制涉及一种持续性低的酶,很可能是病毒逆转录酶。这一观点得到了进一步支持,即形成连接点一部分的大多数CarSV序列要么对应于CarSV RNA所提议构象中的强二级结构基序,要么对应于其自我切割位点,这可能促进了聚合酶跳跃。伴随着最丰富的CarSV RNA,先前已鉴定出一系列具有序列缺失的CarSV RNA。在这里,我们鉴定了它们的一些相应DNA形式,以及迄今在任何已鉴定的CarSV RNA物种中均未发现的其他具有缺失的CarSV DNA形式。其中一些CarSV DNA形式也被检测到与CERV序列融合。这些缩短的CarSV DNA版本的存在可能为其相应转录本提供持续输入,并解释了具有缺陷锤头结构的CarSV RNA的持续存在,对于这种RNA,RNA - RNA扩增模型似乎不太可能。