Zhang Huan, Campbell David A, Sturm Nancy R, Lin Senjie
Department of Marine Sciences, University of Connecticut, CT, USA.
Mol Biol Evol. 2009 Aug;26(8):1757-71. doi: 10.1093/molbev/msp083. Epub 2009 Apr 22.
Spliced leader (SL) trans-splicing is a common mRNA processing mechanism in dinoflagellates, in which a 22-nt sequence is transferred from the 5'-end of a small noncoding RNA, the SL RNA, to the 5'-end of mRNA molecules. Although the SL RNA gene was shown initially to be organized as tandem repeats with transcripts of 50-60 nt, shorter than most of their counterparts in other organisms, other gene organizations and transcript lengths were reported subsequently. To address the evolutionary gradient of gene organization complexity, we thoroughly examined transcript and gene organization of the SL RNA in a phylogenetically and ecologically diverse group of dinoflagellates representing four Orders. All these dinoflagellates possessed SL RNA transcripts of 50-60 nt, although in one species additional transcripts of up to 92 nt were also detected. At the genomic level, various combinations of SL RNA and 5S rRNA tandem gene arrays, including SL RNA-only, 5S rRNA-only, and mixed SL RNA-5S rRNA (SL-5S) clusters, were amplified by polymerase chain reaction for six dinoflagellates, containing intergenic spacers ranging from 88 bp to over 1.2 kb. Of these species, no SL-5S cluster was detected in Prorocentrum minimum, and only Karenia brevis showed the U6 small nuclear RNA gene associated with these mixed arrays. The 5S rRNA-only array was also found in three dinoflagellates, along with two SL-5S-adjacent arrangements found in two other species that could represent junctions. Two species contained multimeric SL exon repeats with no associated intron. These results suggest that 1) both the SL RNA tandem repeat and the SL-5S cluster genomic organizations are an "ancient" and widespread feature within the phylum of dinoflagellates and 2) rampant genomic duplication and recombination are ongoing independently in each dinoflagellate lineage, giving rise to the highly complex and diversified genomic arrangements of the SL RNA gene, while conserving the length and structure of the functional SL RNA.
剪接前导序列(SL)反式剪接是甲藻中一种常见的mRNA加工机制,在此过程中,一个22个核苷酸的序列从小的非编码RNA(SL RNA)的5'端转移到mRNA分子的5'端。尽管最初显示SL RNA基因以50 - 60个核苷酸的转录本串联重复形式存在,比其他生物中的大多数对应物短,但随后报道了其他基因组织形式和转录本长度。为了研究基因组织复杂性的进化梯度,我们全面检查了代表四个目、在系统发育和生态上具有多样性的一组甲藻中SL RNA的转录本和基因组织。所有这些甲藻都拥有50 - 60个核苷酸的SL RNA转录本,尽管在一个物种中还检测到了长达92个核苷酸的额外转录本。在基因组水平上,通过聚合酶链反应对六种甲藻扩增了SL RNA和5S rRNA串联基因阵列的各种组合,包括仅SL RNA、仅5S rRNA以及混合的SL RNA - 5S rRNA(SL - 5S)簇,其基因间隔从88 bp到超过1.2 kb不等。在这些物种中,微小原甲藻未检测到SL - 5S簇,只有短裸甲藻显示出与这些混合阵列相关的U6小核RNA基因。在三种甲藻中还发现了仅5S rRNA阵列,在另外两个物种中发现了两种与SL - 5S相邻的排列,可能代表连接点。两个物种含有多聚体SL外显子重复序列,没有相关内含子。这些结果表明:1)SL RNA串联重复和SL - 5S簇基因组组织形式是甲藻门内“古老”且广泛存在的特征;2)在每个甲藻谱系中,猖獗的基因组复制和重组正在独立进行,导致SL RNA基因的基因组排列高度复杂且多样化,同时保留了功能性SL RNA的长度和结构。