Sharma Niharika, Russell Scott D, Bhalla Prem L, Singh Mohan B
Plant Molecular Biology and Biotechnology Laboratory, Australian Research Council Centre of Excellence for Integrative Legume Research, Melbourne School of Land and Environment, University of Melbourne, Parkville, Victoria 3010, Australia.
BMC Res Notes. 2011 Sep 5;4:319. doi: 10.1186/1756-0500-4-319.
The male germ line in flowering plants is initiated within developing pollen grains via asymmetric division. The smaller cell then becomes totally encased within a much larger vegetative cell, forming a unique "cell within a cell structure". The generative cell subsequently divides to give rise to two non-motile diminutive sperm cells, which take part in double fertilization and lead to the seed set. Sperm cells are difficult to investigate because of their presence within the confines of the larger vegetative cell. However, recently developed techniques for the isolation of rice sperm cells and the fully annotated rice genome sequence have allowed for the characterization of the transcriptional repertoire of sperm cells. Microarray gene expression data has identified a subset of rice genes that show unique or highly preferential expression in sperm cells. This information has led to the identification of cis-regulatory elements (CREs), which are conserved in sperm-expressed genes and are putatively associated with the control of cell-specific expression.
We aimed to identify the CREs associated with rice sperm cell-specific gene expression data using in silico prediction tools. We analyzed 1-kb upstream regions of the top 40 sperm cell co-expressed genes for over-represented conserved and novel motifs. Analysis of upstream regions with the SIGNALSCAN program with the PLACE database, MEME and the Mclip tool helped to find combinatorial sets of known transcriptional factor-binding sites along with two novel motifs putatively associated with the co-expression of sperm cell-specific genes.
Our data shows the occurrence of novel motifs, which are putative CREs and are likely targets of transcriptional factors regulating sperm cell gene expression. These motifs can be used to design the experimental verification of regulatory elements and the identification of transcriptional factors that regulate sperm cell-specific gene expression.
开花植物中的雄性生殖系通过不对称分裂在发育中的花粉粒内起始。较小的细胞随后完全被包裹在一个大得多的营养细胞内,形成一种独特的“细胞中细胞结构”。生殖细胞随后分裂产生两个无运动能力的微小精子细胞,它们参与双受精并导致种子形成。由于精子细胞存在于较大的营养细胞内部,因此很难对其进行研究。然而,最近开发的水稻精子细胞分离技术和完全注释的水稻基因组序列使得对精子细胞转录组的特征分析成为可能。微阵列基因表达数据已经鉴定出水稻基因的一个子集,这些基因在精子细胞中表现出独特或高度优先的表达。这些信息导致了顺式调控元件(CREs)的鉴定,这些元件在精子表达基因中是保守的,并且可能与细胞特异性表达的控制有关。
我们旨在使用计算机预测工具鉴定与水稻精子细胞特异性基因表达数据相关的CREs。我们分析了前40个精子细胞共表达基因的1kb上游区域,以寻找过度代表的保守和新基序。使用PLACE数据库、MEME和Mclip工具通过SIGNALSCAN程序对上游区域进行分析,有助于找到已知转录因子结合位点的组合集以及两个可能与精子细胞特异性基因共表达相关的新基序。
我们的数据显示了新基序的出现,这些基序是假定的CREs,可能是调节精子细胞基因表达的转录因子的靶标。这些基序可用于设计调控元件的实验验证以及鉴定调节精子细胞特异性基因表达的转录因子。