Fischer Cécile, Bouneau Laurence, Coutanceau Jean-Pierre, Weissenbach Jean, Volff Jean-Nicolas, Ozouf-Costaz Catherine
Genoscope/Centre National de Séquençage and CNRS-UMR 8030, F-91057, Evry Cedex 06, France.
Gene. 2004 Jul 21;336(2):175-83. doi: 10.1016/j.gene.2004.04.014.
Because of its unusual high degree of compaction and paucity of repetitive sequences, the genome of the smooth pufferfish Tetraodon nigroviridis is the subject of a well-advanced sequencing project. An astonishing diversity of transposable elements not found in the human and the mouse has been observed in the genome of T. nigroviridis. Due to the difficulty of assembling repeat-rich regions, the whole genome shotgun sequencing approach will probably fail to reveal the general organisation of this compact vertebrate genome. Therefore, in order to gain new insights into the global distribution pattern of repeated DNA in the genome of T. nigroviridis, we have reconstructed partial/complete repetitive sequences from data generated by the genome project and performed double-colour fluorescent in situ hybridization (FISH) analysis for representatives of three major categories of repeated sequences including two minisatellites (ms100 and ms104), two DNA transposons (Tol2 and Buffy1) and two non-long terminal repeat (LTR) retrotransposons (Rex3 and Babar). We show that DNA transposons and retroelements very frequently colocalize with minisatellites and mostly accumulate within heterochromatic regions. These results, which have not been reported so far for the fugu Takifugu rubripes, show that repeated elements are generally excluded from gene-rich regions in T. nigroviridis and underline the extreme degree of compartmentalization of this compact genome. The genome organization of the pufferfish is clearly different from that observed in humans, where repeated sequences make up an important fraction of euchromatic DNA, and is more similar to that observed in the fruit fly Drosophila melanogaster.
由于其异常高的压缩程度和重复序列的稀缺性,光滑河豚黑青斑河豚(Tetraodon nigroviridis)的基因组成为了一个进展良好的测序项目的研究对象。在黑青斑河豚的基因组中观察到了人类和小鼠基因组中未发现的惊人多样的转座元件。由于组装富含重复序列区域的困难,全基因组鸟枪法测序方法可能无法揭示这个紧凑的脊椎动物基因组的总体组织情况。因此,为了深入了解黑青斑河豚基因组中重复DNA的全局分布模式,我们从基因组项目产生的数据中重建了部分/完整的重复序列,并对三类主要重复序列的代表进行了双色荧光原位杂交(FISH)分析,这三类重复序列包括两个微卫星(ms100和ms104)、两个DNA转座子(Tol2和Buffy1)以及两个非长末端重复(LTR)逆转座子(Rex3和Babar)。我们发现DNA转座子和逆转录元件经常与微卫星共定位,并且大多聚集在异染色质区域内。这些结果——迄今尚未在红鳍东方鲀(Takifugu rubripes)中报道——表明在黑青斑河豚中重复元件通常被排除在基因丰富的区域之外,并突显了这个紧凑基因组的极端分隔程度。河豚的基因组组织明显不同于在人类中观察到的情况,在人类中重复序列构成了常染色质DNA的重要部分,并且更类似于在果蝇(Drosophila melanogaster)中观察到的情况。