Ratio-based analysis of differential mRNA processing and expression of a polyadenylation factor mutant pcfs4 using arabidopsis tiling microarray
| Autor: | Yingjia Shen, Denghui Xing, Xiaohui Wu, Jianti Zheng, Qingshun Quinn Li, Diana M. Kroll, Guoli Ji |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
0106 biological sciences
Polyadenylation Microarray Mutant Arabidopsis Computational Biology/Macromolecular Structure Analysis Computational Biology/Transcriptional Regulation lcsh:Medicine Biology Validation Studies as Topic Computational Biology/Molecular Dynamics 01 natural sciences Genome Genetics and Genomics/Plant Genetics and Gene Expression Molecular Biology/Bioinformatics 03 medical and health sciences Gene Expression Regulation Plant Computational Biology/Alternative Splicing RNA Messenger RNA Processing Post-Transcriptional lcsh:Science 030304 developmental biology Genetics mRNA Cleavage and Polyadenylation Factors 0303 health sciences Messenger RNA Multidisciplinary Rna processing Arabidopsis Proteins Genetics and Genomics/Functional Genomics Gene Expression Profiling lcsh:R Genetics and Genomics/Bioinformatics Reference Standards biology.organism_classification Microarray Analysis Plants Genetically Modified Developmental Biology/Plant Growth and Development Computational Biology/Sequence Motif Analysis Mutant Proteins lcsh:Q DNA microarray Algorithms 010606 plant biology & botany Research Article |
| Zdroj: | PLoS ONE, Vol 6, Iss 2, p e14719 (2011) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Background Alternative polyadenylation as a mechanism in gene expression regulation has been widely recognized in recent years. Arabidopsis polyadenylation factor PCFS4 was shown to function in leaf development and in flowering time control. The function of PCFS4 in controlling flowering time was correlated with the alternative polyadenylation of FCA, a flowering time regulator. However, genetic evidence suggested additional targets of PCFS4 that may mediate its function in both flowering time and leaf development. Methodology/Principal Findings To identify further targets, we investigated the whole transcriptome of a PCFS4 mutant using Affymetrix Arabidopsis genomic tiling 1.0R array and developed a data analysis pipeline, termed RADPRE (Ratio-based Analysis of Differential mRNA Processing and Expression). In RADPRE, ratios of normalized probe intensities between wild type Columbia and a pcfs4 mutant were first generated. By doing so, one of the major problems of tiling array data—variations caused by differential probe affinity—was significantly alleviated. With the probe ratios as inputs, a hierarchy of statistical tests was carried out to identify differentially processed genes (DPG) and differentially expressed genes (DEG). The false discovery rate (FDR) of this analysis was estimated by using the balanced random combinations of Col/pcfs4 and pcfs4/Col ratios as inputs. Gene Ontology (GO) analysis of the DPGs and DEGs revealed potential new roles of PCFS4 in stress responses besides flowering time regulation. Conclusion/Significance We identified 68 DPGs and 114 DEGs with FDR at 1% and 2%, respectively. Most of the 68 DPGs were subjected to alternative polyadenylation, splicing or transcription initiation. Quantitative PCR analysis of a set of DPGs confirmed that most of these genes were truly differentially processed in pcfs4 mutant plants. The enriched GO term “regulation of flower development” among PCFS4 targets further indicated the efficacy of the RADPRE pipeline. This simple but effective program is available upon request. |
| Databáze: | OpenAIRE |
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