拟南芥和水稻锚蛋白（ANKP）基因家族初步研究
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摘要
锚蛋白是一类十分重要的结联蛋白家族，其结构的主要特征是含有数量不一的锚定重复序列模体，主要作用是介导蛋白质与蛋白质的相互作用，广泛参与细胞生命活动的调节。本论文在拟南芥和水稻基因组测序工作完成的基础上，利用生物信息学的方法，鉴定和分析了两种模式植物基因组的锚定蛋白基因。研究结果如下：
1．鉴定得到108个拟南芥基因组锚定蛋白基因，并根据蛋白质二级结构分析，将它们进行聚类分析，共分成3个大组10个小组。拟南芥基因芯片数据库的查询和分析，得到一些植物特有的锚定蛋白基因，主要表现在对植物病原体、逆境、光节律以及蛋白质合成等方面，但它们的功能需要实验进一步确定。
2．基于基因芯片的分析结果，选择10个拟抗病相关基因，在转录水平上初步研究了拟南芥拟抗性基因受植物病原体诱导后的表达情况，发现At1g10340对镰刀菌(Fusarium oxysporum)敏感，在其侵染后的第2天出现表达高峰。
3．克隆和初步分析了拟南芥基因At1g10340，基因表达模式分析表明At1g10340在除角果外均有表达，亚细胞定位分析发现At1g10340定位于质膜上。
4．鉴定得到142个水稻基因组锚蛋白基因，并根据蛋白质二级结构分析，将它们进行聚类分析，共分成3个大组14个小组。并用RT-PCR的方法分析了水稻锚定膜蛋白基因ANKTM的表达模式。
5．比较和分析了拟南芥和水稻基因组的锚蛋白基因。首先，跟动物或其它生物比较，有些锚蛋白基因在各生物体比较保守，这些保守基因可能存在于所有生物中，作为细胞生活的基本组成部分。其次，两种模式植物的锚蛋白基因比较表明，它们在基因组水平存在一定程度的相似性，可能代表着植物所共有的生理功能。同时，两者之间也存在一定的差异，这些植物锚蛋白分子可能是单子叶或双子叶植物特有的，或者为适应陆生或水生等不同的生活环境而不断进化发展而形成的。
ANK protein is an important member of the binding protein family, which containing ankyrin repeats motif, mediated in protein-protein recognition, may involved in the regulation of diverse cellular activity. Based on genome sequencing of Arabidopsis and rice, this paper identified and analyzed the ANK genes of two model plants using the methodes of bioinformatics. The results as follows:1. There are 108 ANK genes of arabidopsis which have been identified. Acoording to cluster analysis of secondary structures of the putative proteins, these genes can be mainly classified into three large groups which can further be divided into 14 subgroups. Microarray data of arabidopsis indicate that some of these genes can only be found in plants, which may play roles in protein synthesis, pathogenic responses, stress, light rhythm and so on. But the function of these genes need to be further proved.2. Fusarium-induced pathogenic responses of the putative arabidopsis resistant genes were studied by testing the transcriptional expression level of the genes. At1g10340 has been proved to be sensitive to Fusarium oxysporum.3. cDNA of At1g10340, was obtained by RT-PCR. Transcriptional gene expression pattern showed it expressed in all tissues except for silique, Subcellular localisation analysis showed it located on plasma membrane.
4. In rice, 142 ANKP genes have been identified, which can be classified into three large groups which divided into 14 subgroups by the same way of cluster analysis in arabidopsis. And the expression pattern of the rice ANKTM genes were analyzed by RT-PCR.
5. Compared and analysized ANKP genes in rice and arabidopsis genome. First, Compared with other organisms, some of ANKP genes are conserved across eukaryotic organisms, they may act in basic cellular activities. Second, compared between rice and arabidopsis, they share similarity to some extent in view of the level of genome. The similarity of plant specific ANKP genes in the two kinds plants suggested that they may play roles in all kinds of plants. Tthe divergence of plant specific ANKP genes between these two model plants suggested that these ANKP genes may evolved toward dicot specific or monocot specific respectively, or they involved to adapt terrestrial or hydrophytic inhabitats.
1．鉴定得到108个拟南芥基因组锚定蛋白基因，并根据蛋白质二级结构分析，将它们进行聚类分析，共分成3个大组10个小组。拟南芥基因芯片数据库的查询和分析，得到一些植物特有的锚定蛋白基因，主要表现在对植物病原体、逆境、光节律以及蛋白质合成等方面，但它们的功能需要实验进一步确定。
2．基于基因芯片的分析结果，选择10个拟抗病相关基因，在转录水平上初步研究了拟南芥拟抗性基因受植物病原体诱导后的表达情况，发现At1g10340对镰刀菌(Fusarium oxysporum)敏感，在其侵染后的第2天出现表达高峰。
3．克隆和初步分析了拟南芥基因At1g10340，基因表达模式分析表明At1g10340在除角果外均有表达，亚细胞定位分析发现At1g10340定位于质膜上。
4．鉴定得到142个水稻基因组锚蛋白基因，并根据蛋白质二级结构分析，将它们进行聚类分析，共分成3个大组14个小组。并用RT-PCR的方法分析了水稻锚定膜蛋白基因ANKTM的表达模式。
5．比较和分析了拟南芥和水稻基因组的锚蛋白基因。首先，跟动物或其它生物比较，有些锚蛋白基因在各生物体比较保守，这些保守基因可能存在于所有生物中，作为细胞生活的基本组成部分。其次，两种模式植物的锚蛋白基因比较表明，它们在基因组水平存在一定程度的相似性，可能代表着植物所共有的生理功能。同时，两者之间也存在一定的差异，这些植物锚蛋白分子可能是单子叶或双子叶植物特有的，或者为适应陆生或水生等不同的生活环境而不断进化发展而形成的。
ANK protein is an important member of the binding protein family, which containing ankyrin repeats motif, mediated in protein-protein recognition, may involved in the regulation of diverse cellular activity. Based on genome sequencing of Arabidopsis and rice, this paper identified and analyzed the ANK genes of two model plants using the methodes of bioinformatics. The results as follows:1. There are 108 ANK genes of arabidopsis which have been identified. Acoording to cluster analysis of secondary structures of the putative proteins, these genes can be mainly classified into three large groups which can further be divided into 14 subgroups. Microarray data of arabidopsis indicate that some of these genes can only be found in plants, which may play roles in protein synthesis, pathogenic responses, stress, light rhythm and so on. But the function of these genes need to be further proved.2. Fusarium-induced pathogenic responses of the putative arabidopsis resistant genes were studied by testing the transcriptional expression level of the genes. At1g10340 has been proved to be sensitive to Fusarium oxysporum.3. cDNA of At1g10340, was obtained by RT-PCR. Transcriptional gene expression pattern showed it expressed in all tissues except for silique, Subcellular localisation analysis showed it located on plasma membrane.
4. In rice, 142 ANKP genes have been identified, which can be classified into three large groups which divided into 14 subgroups by the same way of cluster analysis in arabidopsis. And the expression pattern of the rice ANKTM genes were analyzed by RT-PCR.
5. Compared and analysized ANKP genes in rice and arabidopsis genome. First, Compared with other organisms, some of ANKP genes are conserved across eukaryotic organisms, they may act in basic cellular activities. Second, compared between rice and arabidopsis, they share similarity to some extent in view of the level of genome. The similarity of plant specific ANKP genes in the two kinds plants suggested that they may play roles in all kinds of plants. Tthe divergence of plant specific ANKP genes between these two model plants suggested that these ANKP genes may evolved toward dicot specific or monocot specific respectively, or they involved to adapt terrestrial or hydrophytic inhabitats.
引文
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[3] 杜海宁，胡红雨．锚蛋白重复序列介导的蛋白质．蛋白质相互作用[J]．生物化学与生物物理进展，2002，29(1)：6-9。
[4] Mosavi LK, Minor DL Jr, Peng ZY. Consensus-derived structural determinants of the ankyrin repeat motif [J]. Proc Natl Aead Sci U S A, 2002, 99(25): 16029-16034.
[5] Rupee RA, Poujol D, Grosgeorge J, Carle GF, Livolsi A, Peyron JF, Schmid RM, Baeuerle PA, Messer G.Structural analysis, expression, and chromosomal localization of the mouse ikba gene [J]. Immunogenetics. 1999 49(5):395-403.
[6] Shimada M, Satoh N, Yokosawa H.Involvement of Rel/NF-kappaB in regulation of ascidian notoehord formation [J]. Dev Growth Differ. 2001,43(2): 145-54.
[7] Mosavi LK, Cammett TJ, Desrosiers DC, Peng ZY. The ankyrin repeat as molecular architecturefor protein recognition [J].Protein Science, 2004, 13(6):1435-1448.
[8] Russo AA, Tong L, Pavletich NP, et al. Structural basis for inhibition of the cyelin-dependent kinase Cdk6 by the tumour suppressor p16INK4a [J]. Nature, 1998, 395(6699):237-143.
[9] Mark EZ, Daniel JL, Frederick MH. Structure and stability of the ankyrin domain of the Drosophila Notch receptor [J]. Protein Science, 2003, 12(11):2622-2632.
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