Dufresne Marie, Hua-Van Aurélie, El Wahab Hala Abd, Ben M'Barek Sarrah, Vasnier Christelle, Teysset Laure, Kema Gert H J, Daboussi Marie-Josée
Institut de Génétique et Microbiologie, Université Paris-Sud, UMR8621, F-91405 Orsay, France.
Genetics. 2007 Jan;175(1):441-52. doi: 10.1534/genetics.106.064360. Epub 2006 Dec 18.
The mimp1 element previously identified in the ascomycete fungus Fusarium oxysporum has hallmarks of miniature inverted-repeat transposable elements (MITEs): short size, terminal inverted repeats (TIRs), structural homogeneity, and a stable secondary structure. Since mimp1 has no coding capacity, its mobilization requires a transposase-encoding element. On the basis of the similarity of TIRs and target-site preference with the autonomous Tc1-like element impala, together with a correlated distribution of both elements among the Fusarium genus, we investigated the ability of mimp1 to jump upon expression of the impala transposase provided in trans. Under these conditions, we present evidence that mimp1 transposes by a cut-and-paste mechanism into TA dinucleotides, which are duplicated upon insertion. Our results also show that mimp1 reinserts very frequently in genic regions for at least one-third of the cases. We also show that the mimp1/impala double-component system is fully functional in the heterologous species F. graminearum, allowing the development of a highly efficient tool for gene tagging in filamentous fungi.
先前在子囊菌镰刀菌中鉴定出的mimp1元件具有微型反向重复转座元件(MITEs)的特征:尺寸短小、末端反向重复序列(TIRs)、结构同源性以及稳定的二级结构。由于mimp1没有编码能力,其转座需要一个编码转座酶的元件。基于TIRs与自主的类Tc1元件黑斑羚(impala)的相似性以及靶位点偏好性,再加上这两种元件在镰刀菌属中的相关分布,我们研究了在反式提供黑斑羚转座酶表达时mimp1跳跃的能力。在这些条件下,我们提供了证据表明mimp1通过剪切粘贴机制转座到TA二核苷酸中,插入时TA二核苷酸会发生重复。我们的结果还表明,在至少三分之一的情况下,mimp1非常频繁地重新插入基因区域。我们还表明,mimp1/黑斑羚双组分系统在异源物种禾谷镰刀菌中完全功能正常,从而为丝状真菌中的基因标签开发了一种高效工具。