Lakshmanan Vairavan, Bansal Dhiru, Kulkarni Jahnavi, Poduval Deepak, Krishna Srikar, Sasidharan Vidyanand, Anand Praveen, Seshasayee Aswin, Palakodeti Dasaradhi
Institute for Stem Cell Biology and Regenerative Medicine, NCBS campus, GKVK, Bellary Road, Bangalore 560065, Karnataka, India SASTRA University, Thirumalaisamudram, Thanjavur 613401, Tamil Nadu, India.
Institute for Stem Cell Biology and Regenerative Medicine, NCBS campus, GKVK, Bellary Road, Bangalore 560065, Karnataka, India Manipal University, Madhav Nagar, Near Tiger Circle, Manipal 576104, Karnataka, India.
G3 (Bethesda). 2016 Oct 13;6(10):3035-3048. doi: 10.1534/g3.116.031120.
In eukaryotes, 3' untranslated regions (UTRs) play important roles in regulating posttranscriptional gene expression. The 3'UTR is defined by regulated cleavage/polyadenylation of the pre-mRNA. The advent of next-generation sequencing technology has now enabled us to identify these events on a genome-wide scale. In this study, we used poly(A)-position profiling by sequencing (3P-Seq) to capture all poly(A) sites across the genome of the freshwater planarian, Schmidtea mediterranea, an ideal model system for exploring the process of regeneration and stem cell function. We identified the 3'UTRs for ∼14,000 transcripts and thus improved the existing gene annotations. We found 97 transcripts, which are polyadenylated within an internal exon, resulting in the shrinking of the ORF and loss of a predicted protein domain. Around 40% of the transcripts in planaria were alternatively polyadenylated (ApA), resulting either in an altered 3'UTR or a change in coding sequence. We identified specific ApA transcript isoforms that were subjected to miRNA mediated gene regulation using degradome sequencing. In this study, we also confirmed a tissue-specific expression pattern for alternate polyadenylated transcripts. The insights from this study highlight the potential role of ApA in regulating the gene expression essential for planarian regeneration.
在真核生物中,3'非翻译区(3'UTR)在转录后基因表达调控中发挥着重要作用。3'UTR由前体mRNA的调控性切割/聚腺苷酸化所定义。新一代测序技术的出现使我们能够在全基因组范围内识别这些事件。在本研究中,我们使用测序法进行聚腺苷酸位点分析(3P-Seq)来捕获淡水涡虫地中海实蝇(Schmidtea mediterranea)全基因组中的所有聚腺苷酸位点,这是探索再生过程和干细胞功能的理想模型系统。我们确定了约14,000个转录本的3'UTR,从而改进了现有的基因注释。我们发现97个转录本在内部外显子内进行聚腺苷酸化,导致开放阅读框缩小和预测的蛋白质结构域丢失。涡虫中约40%的转录本存在可变聚腺苷酸化(ApA),这要么导致3'UTR改变,要么导致编码序列变化。我们使用降解组测序鉴定了受miRNA介导的基因调控的特定ApA转录本异构体。在本研究中,我们还证实了可变聚腺苷酸化转录本的组织特异性表达模式。这项研究的见解突出了ApA在调控涡虫再生所必需的基因表达中的潜在作用。