Rubin E, Levy A A
Department of Plant Genetics, The Weizmann Institute of Science, Rehovot, Israel.
Mol Cell Biol. 1997 Nov;17(11):6294-302. doi: 10.1128/MCB.17.11.6294.
The mechanism by which the maize autonomous Ac transposable element gives rise to nonautonomous Ds elements is largely unknown. Sequence analysis of native maize Ds elements indicates a complex chimeric structure formed through deletions of Ac sequences with or without insertions of Ac-unrelated sequence blocks. These blocks are often flanked by short stretches of reshuffled and duplicated Ac sequences. To better understand the mechanism leading to Ds formation, we designed an assay for detecting alterations in Ac using transgenic tobacco plants carrying a single copy of Ac. We found frequent de novo alterations in Ac which were excision rather than sequence dependent, occurring within Ac but not within an almost identical Ds element and not within a stable transposase-producing gene. The de novo DNA rearrangements consisted of internal deletions with breakpoints usually occurring at short repeats and, in some cases, of duplication of Ac sequences or insertion of Ac-unrelated fragments. The ancient maize Ds elements and the young Ds elements in transgenic tobacco showed similar rearrangements, suggesting that Ac-Ds elements evolve rapidly, more so than stable genes, through deletions, duplications, and reshuffling of their own sequences and through capturing of unrelated sequences. The data presented here suggest that abortive Ac-induced gap repair, through the synthesis-dependent strand-annealing pathway, is the underlying mechanism for Ds element formation.
玉米自主型Ac转座元件产生非自主型Ds元件的机制在很大程度上尚不清楚。对天然玉米Ds元件的序列分析表明,其形成了一种复杂的嵌合结构,该结构是通过缺失Ac序列(有无插入与Ac无关的序列块)而形成的。这些序列块通常两侧是短片段的重新排列和重复的Ac序列。为了更好地理解导致Ds形成的机制,我们设计了一种检测方法,利用携带单个Ac拷贝的转基因烟草植株来检测Ac的变化。我们发现Ac中频繁出现从头变化,这些变化是切除而非序列依赖性的,发生在Ac内部,但不在几乎相同的Ds元件内部,也不在稳定的转座酶产生基因内部。从头DNA重排包括内部缺失,断点通常出现在短重复序列处,在某些情况下还包括Ac序列的重复或插入与Ac无关的片段。转基因烟草中的古代玉米Ds元件和年轻Ds元件表现出相似的重排,这表明Ac-Ds元件通过自身序列的缺失、重复和重排以及捕获无关序列,比稳定基因进化得更快。本文提供的数据表明,通过合成依赖链退火途径的Ac诱导的无效缺口修复是Ds元件形成的潜在机制。