Minic Zoran, Jamet Elisabeth, San-Clemente Hélène, Pelletier Sandra, Renou Jean-Pierre, Rihouey Christophe, Okinyo Denis P O, Proux Caroline, Lerouge Patrice, Jouanin Lise
Department of Chemistry, University of Saskatchewan, 110 Science Place, Saskatoon, SK, Canada.
BMC Plant Biol. 2009 Jan 16;9:6. doi: 10.1186/1471-2229-9-6.
Different strategies (genetics, biochemistry, and proteomics) can be used to study proteins involved in cell biogenesis. The availability of the complete sequences of several plant genomes allowed the development of transcriptomic studies. Although the expression patterns of some Arabidopsis thaliana genes involved in cell wall biogenesis were identified at different physiological stages, detailed microarray analysis of plant cell wall genes has not been performed on any plant tissues. Using transcriptomic and bioinformatic tools, we studied the regulation of cell wall genes in Arabidopsis stems, i.e. genes encoding proteins involved in cell wall biogenesis and genes encoding secreted proteins.
Transcriptomic analyses of stems were performed at three different developmental stages, i.e., young stems, intermediate stage, and mature stems. Many genes involved in the synthesis of cell wall components such as polysaccharides and monolignols were identified. A total of 345 genes encoding predicted secreted proteins with moderate or high level of transcripts were analyzed in details. The encoded proteins were distributed into 8 classes, based on the presence of predicted functional domains. Proteins acting on carbohydrates and proteins of unknown function constituted the two most abundant classes. Other proteins were proteases, oxido-reductases, proteins with interacting domains, proteins involved in signalling, and structural proteins. Particularly high levels of expression were established for genes encoding pectin methylesterases, germin-like proteins, arabinogalactan proteins, fasciclin-like arabinogalactan proteins, and structural proteins. Finally, the results of this transcriptomic analyses were compared with those obtained through a cell wall proteomic analysis from the same material. Only a small proportion of genes identified by previous proteomic analyses were identified by transcriptomics. Conversely, only a few proteins encoded by genes having moderate or high level of transcripts were identified by proteomics.
Analysis of the genes predicted to encode cell wall proteins revealed that about 345 genes had moderate or high levels of transcripts. Among them, we identified many new genes possibly involved in cell wall biogenesis. The discrepancies observed between results of this transcriptomic study and a previous proteomic study on the same material revealed post-transcriptional mechanisms of regulation of expression of genes encoding cell wall proteins.
可采用不同策略(遗传学、生物化学和蛋白质组学)来研究参与细胞生物合成的蛋白质。几种植物基因组完整序列的可得性推动了转录组学研究的开展。尽管已在不同生理阶段鉴定了一些拟南芥中参与细胞壁生物合成的基因的表达模式,但尚未对任何植物组织进行植物细胞壁基因的详细微阵列分析。我们利用转录组学和生物信息学工具,研究了拟南芥茎中细胞壁基因的调控情况,即参与细胞壁生物合成的蛋白质编码基因和分泌蛋白编码基因。
在三个不同发育阶段对茎进行了转录组分析,即幼茎、中间阶段和成熟茎。鉴定出了许多参与细胞壁成分(如多糖和单木质醇)合成的基因。对总共345个编码预测分泌蛋白且转录本水平中等或较高的基因进行了详细分析。根据预测功能域的存在情况,将编码的蛋白质分为8类。作用于碳水化合物的蛋白质和功能未知的蛋白质构成了最丰富的两类。其他蛋白质包括蛋白酶、氧化还原酶、具有相互作用结构域的蛋白质、参与信号传导的蛋白质和结构蛋白。果胶甲酯酶、类萌发素蛋白、阿拉伯半乳聚糖蛋白、类成束蛋白阿拉伯半乳聚糖蛋白和结构蛋白编码基因的表达水平特别高。最后,将该转录组分析的结果与通过对相同材料进行细胞壁蛋白质组分析获得的结果进行了比较。先前蛋白质组分析鉴定的基因中只有一小部分通过转录组学得以鉴定。相反,蛋白质组学仅鉴定出了转录本水平中等或较高的基因所编码的少数蛋白质。
对预测编码细胞壁蛋白的基因进行分析发现,约345个基因具有中等或较高水平的转录本。其中,我们鉴定出了许多可能参与细胞壁生物合成的新基因。该转录组研究结果与先前对相同材料进行的蛋白质组研究结果之间的差异揭示了细胞壁蛋白编码基因表达的转录后调控机制。