Pennington R E, Melcher U
Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater 74078-0454.
Virology. 1993 Jan;192(1):188-96. doi: 10.1006/viro.1993.1021.
RNA splicing and copy-choice recombination lead to deletions in the DNA of cauliflower mosaic virus (CaMV). To assess the relative importance of these mechanisms of nucleotide sequence change, free of constraints imposed by the "relay race" mode of translation of CaMV RNA, we examined the stability of inserts in the large intergenic region. The insertions had, in various combinations, splicing signals and directly repeated sequences that could facilitate deletion. Most modified DNAs were infectious. Viral DNA recovered from infected plants was analyzed by restriction and, in some cases, cloned for nucleotide sequencing to determine deletion endpoints. Deletions from a DNA containing introduced splicing signals occurred primarily at direct repeats, although deletion apparently by splicing was also detected. Both types of deletion were also observed with insertions containing a 5' splice donor but no known functional 3' splice acceptor. One DNA, whose insertion lacked splicing signals and short repeated sequences, was stable in one of two plants infected. Total insert deletions were bounded by repeats or pseudorepeats, while partial insert deletions apparently occurred by splicing. The results suggest that the two mechanisms for deletion of nucleotides are equally important in the evolution of caulimoviral nucleotide sequences.
RNA剪接和模板转换重组导致花椰菜花叶病毒(CaMV)DNA发生缺失。为了评估这些核苷酸序列变化机制的相对重要性,不受CaMV RNA“接力赛”翻译模式所施加的限制,我们检测了大基因间隔区插入片段的稳定性。这些插入片段以各种组合形式含有剪接信号和可能促进缺失的直接重复序列。大多数修饰后的DNA具有感染性。从受感染植物中回收的病毒DNA通过限制性内切酶分析,在某些情况下,进行克隆以进行核苷酸测序,以确定缺失的端点。含有引入的剪接信号的DNA中的缺失主要发生在直接重复序列处,不过也检测到了明显由剪接导致的缺失。在含有5'剪接供体但没有已知功能性3'剪接受体的插入片段中也观察到了这两种类型的缺失。一个插入片段缺乏剪接信号和短重复序列的DNA在两株受感染植物中的一株中是稳定的。总插入片段缺失由重复序列或假重复序列界定,而部分插入片段缺失显然是由剪接导致的。结果表明,这两种核苷酸缺失机制在花椰菜花叶病毒核苷酸序列的进化中同样重要。