Teodorovic Smilja, Walls Colleen D, Elmendorf Heidi G
Biology Department, 406 Reiss Bldg., 37th and O Sts. NW, Georgetown University, Washington DC 20057, USA.
Nucleic Acids Res. 2007;35(8):2544-53. doi: 10.1093/nar/gkm105. Epub 2007 Apr 1.
A prominent feature of transcription in Giardia lamblia is the abundant production of sterile antisense transcripts (Elmendorf et al. The abundance of sterile transcripts in Giardia lamblia. Nucleic Acids., 29, 4674-4683). Here, we use a computational biology analysis of SAGE data to assess the abundance and distribution of sense and antisense messages in the parasite genome. Sterile antisense transcripts are produced at approximately 50% of loci with detectable transcription, yet their abundance at a given locus does not correlate to the abundance of the complementary sense transcripts at that locus or to transcription levels at neighboring loci. These data suggest that sterile antisense transcripts are not simply a local effect of open chromatin structure. Using 5'RACE, we demonstrate that Giardia promoters are a source of antisense transcripts through bidirectional transcription, producing both downstream coding sense and upstream sterile antisense transcripts. We use a dual reporter system to explore roles of specific promoter elements in this bidirectional initiation of transcription and suggest that the degenerate AT-rich nature of TATA and Inr elements in Giardia permits them to function interchangeably. The phenomenon of bidirectional transcription in G. lamblia gives us insight into the interaction between transcriptional machinery and promoter elements, and may be the prominent source of the abundant antisense transcription in this parasite.
蓝氏贾第鞭毛虫转录的一个显著特征是大量产生无义反义转录本(埃尔门多夫等人,《蓝氏贾第鞭毛虫中无义转录本的丰度》,《核酸研究》,第29卷,第4674 - 4683页)。在此,我们利用对SAGE数据的计算生物学分析来评估该寄生虫基因组中正义与反义信息的丰度和分布。在约50%可检测到转录的基因座处产生无义反义转录本,然而它们在给定基因座处的丰度与该基因座处互补正义转录本的丰度或相邻基因座的转录水平均无关联。这些数据表明无义反义转录本并非仅仅是开放染色质结构的局部效应。利用5'RACE技术,我们证明蓝氏贾第鞭毛虫启动子是通过双向转录产生反义转录本的来源,既产生下游编码正义转录本,也产生上游无义反义转录本。我们使用双报告系统来探究特定启动子元件在这种双向转录起始中的作用,并表明蓝氏贾第鞭毛虫中TATA和Inr元件富含AT的简并性质使它们能够互换功能。蓝氏贾第鞭毛虫中的双向转录现象让我们深入了解了转录机制与启动子元件之间的相互作用,并且可能是该寄生虫中大量反义转录的主要来源。