Huang Peiming, Pleasance Erin D, Maydan Jason S, Hunt-Newbury Rebecca, O'Neil Nigel J, Mah Allan, Baillie David L, Marra Marco A, Moerman Donald G, Jones Steven J M
Genome Sciences Centre, BC Cancer Agency, Vancouver, British Columbia V5Z 1L3, Canada.
Genome Res. 2007 Oct;17(10):1478-85. doi: 10.1101/gr.6824707. Epub 2007 Aug 21.
The current Caenorhabditis elegans genomic annotation has many genes organized in operons. Using directionally stitched promoterGFP methodology, we have conducted the largest survey to date on the regulatory regions of annotated C. elegans operons and identified 65, over 25% of those studied, with internal promoters. We have termed these operons "hybrid operons." GFP expression patterns driven from internal promoters differ in tissue specificity from expression of operon promoters, and serial analysis of gene expression data reveals that there is a lack of expression correlation between genes in many hybrid operons. The average length of intergenic regions with putative promoter activity in hybrid operons is larger than previous estimates for operons as a whole. Genes with internal promoters are more commonly involved in gene duplications and have a significantly lower incidence of alternative splicing than genes without internal promoters, although we have observed almost all trans-splicing patterns in these two distinct groups. Finally, internal promoter constructs are able to rescue lethal knockout phenotypes, demonstrating their necessity in gene regulation and survival. Our work suggests that hybrid operons are common in the C. elegans genome and that internal promoters influence not only gene organization and expression but also operon evolution.
当前的秀丽隐杆线虫基因组注释中有许多基因以操纵子形式组织。使用定向拼接启动子绿色荧光蛋白(promoterGFP)方法,我们对注释的秀丽隐杆线虫操纵子的调控区域进行了迄今为止最大规模的调查,并鉴定出65个具有内部启动子的操纵子,占所研究操纵子的25%以上。我们将这些操纵子称为“杂交操纵子”。由内部启动子驱动的绿色荧光蛋白表达模式在组织特异性上与操纵子启动子的表达不同,并且基因表达数据的序列分析表明,许多杂交操纵子中的基因之间缺乏表达相关性。杂交操纵子中具有假定启动子活性的基因间区域的平均长度大于之前对整个操纵子的估计。具有内部启动子的基因更常参与基因复制,并且与没有内部启动子的基因相比,可变剪接的发生率显著更低,尽管我们在这两个不同的组中观察到了几乎所有的反式剪接模式。最后,内部启动子构建体能够挽救致死性敲除表型,证明了它们在基因调控和生存中的必要性。我们的工作表明杂交操纵子在秀丽隐杆线虫基因组中很常见,并且内部启动子不仅影响基因组织和表达,还影响操纵子进化。