Kamikawa Ryoma, Inagaki Yuji, Hashimoto Tetsuo
Graduate School of Human and Environmental Studies, Kyoto University, Yoshida-Nihonmatsu cho, Kyoto 606-8501, Japan.
BMC Res Notes. 2014 Jun 30;7:413. doi: 10.1186/1756-0500-7-413.
Giardia intestinalis is a parasitic unicellular eukaryote with a highly reduced genome, in which only six cis-spliced and four trans-spliced introns have been discovered. However, we anticipate that more cis- and trans-spliced introns likely remain unidentified in genes encoding hypothetical proteins that occupy ca. 2/3 of all of the open reading frames (ORFs) in the Giardia genome. Consequently, comprehensive surveys of introns in ORFs for hypothetical proteins are critical for better understanding of the intron evolution in this organism.
In this study, we identified two novel cis-spliced introns in the draft genome data of G. intestinalis strain WB, by surveying the conserved sequence motifs shared amongst the previously known introns. G. intestinalis strains can be divided into phylogenetically distinct assemblages A-H, and all the introns identified in past studies are shared among the published genome data from strains WB, DH, GS, and P15 representing assemblages A1, A2, B, and E, respectively. Nevertheless one of the two novel introns identified in this study was found to be absent in strain P15.
By considering the organismal relationship among G. intestinalis assemblages A1, A2, B, and E, one of the two introns identified in this study has highly likely been lost after the divergence of the assemblages. On the basis of a sequence comparison between the intron-bearing loci in WB, DH, and GS genomes and the homologous but intron-free locus in P15 genome, we propose that the loss of this particular intron was mediated by integration of the DNA fragment reverse-transcribed from mature mRNAs.
肠贾第虫是一种寄生性单细胞真核生物,其基因组高度简化,仅发现了六个顺式剪接内含子和四个反式剪接内含子。然而,我们预计在编码假定蛋白的基因中可能仍有更多顺式和反式剪接内含子未被识别,这些假定蛋白约占贾第虫基因组中所有开放阅读框(ORF)的2/3。因此,全面调查假定蛋白ORF中的内含子对于更好地理解该生物体中内含子的进化至关重要。
在本研究中,我们通过调查先前已知内含子之间共享的保守序列基序,在肠贾第虫WB株的基因组草图数据中鉴定出两个新的顺式剪接内含子。肠贾第虫菌株可分为系统发育上不同的A - H组,过去研究中鉴定出的所有内含子在分别代表A1、A2、B和E组的WB、DH、GS和P15株的已发表基因组数据中均有共享。然而,本研究中鉴定出的两个新内含子之一在P15株中未被发现。
考虑到肠贾第虫A1、A2、B和E组之间的生物关系,本研究中鉴定出的两个内含子之一很可能在这些组分化后丢失。基于WB、DH和GS基因组中含内含子位点与P15基因组中同源但无内含子位点之间的序列比较,我们提出该特定内含子的丢失是由成熟mRNA逆转录的DNA片段整合介导的。