Waksman Institute, Rutgers University, Piscataway, New Jersey 08854, USA.
Plant Cell. 2012 Oct;24(10):4149-62. doi: 10.1105/tpc.112.104265. Epub 2012 Oct 30.
The autonomous transposon Activator (Ac) is a powerful mutagen. Ac-induced mutations range from small footprints of host sequences to large rearrangements of transposon or host sequences. These mutations arise by different repair mechanisms of the double-strand break produced by Ac excision: footprints by nonhomologous end joining and rearrangements by various mechanisms, including DNA replication repair. Footprints greatly outnumber other mutations, masking them because they usually share a nonfunctional phenotype. To determine the spectrum and frequencies of host and self-mutations generated by Ac, we used an allele harboring Ac in the 5' untranslated region bronze (bz). In this system, simple excisions produce purple revertants, whereas deletions of host or transposon sequences produce stable bronze (bz-s) mutants. Internal and terminal deletions of Ac predominated among the 72 bz-s derivatives. Most internal deletions (52 of 54) behaved as nonautonomous Dissociation (Ds) elements. All nine terminal deletions or fractured Ac (fAc) elements had rearrangements of adjacent host sequences. Most Ds and fAc deletion junctions displayed microhomologies and contained filler DNA from nearby sequences, suggesting an origin by DNA repair synthesis followed by microhomology-mediated end joining. All mutations occurred more frequently in pollen, where one in 200 grains carried new Ds or fAc elements.
自主转座子激活子 (Ac) 是一种强大的诱变剂。Ac 诱导的突变范围从小的宿主序列足迹到转座子或宿主序列的大重排。这些突变是由 Ac 切除产生的双链断裂的不同修复机制引起的:足迹通过非同源末端连接,而重排则通过各种机制,包括 DNA 复制修复。足迹的数量大大超过其他突变,掩盖了它们,因为它们通常具有非功能表型。为了确定 Ac 产生的宿主和自身突变的谱和频率,我们使用了一个在 5'非翻译区含有 Ac 的等位基因 bronze(bz)。在这个系统中,简单的切除产生紫色回复突变体,而宿主或转座子序列的缺失则产生稳定的 bronze(bz-s)突变体。Ac 的内部和末端缺失在 72 个 bz-s 衍生物中占主导地位。大多数内部缺失 (54 个中的 52 个) 表现为非自主解离 (Ds) 元件。所有九个末端缺失或断裂的 Ac(fAc) 元件都有相邻宿主序列的重排。大多数 Ds 和 fAc 缺失连接处显示微同源性,并包含来自附近序列的填充 DNA,表明起源于 DNA 修复合成,随后是微同源介导的末端连接。所有突变在花粉中更频繁发生,每 200 个花粉中有一个携带新的 Ds 或 fAc 元件。